PRIORITY
[0001] This application claims the benefit of Provisional Application Serial number
61/258,914 and Provisional Application Serial number
61/258,918 that were both filed on November 6, 2009.
FIELD OF THE DISCLOSURE
[0002] Disclosed herein are prolyl hydroxylase inhibitors that can stabilize hypoxia inducible
factor-1 alpha (HIF-1α), as well as hypoxia inducible factor-2 alpha (HIF-2α). Also
disclosed herein are pharmaceutical compositions comprising one or more of the disclosed
compounds. Yet further disclosed are methods for stimulating the cellular immune response
in a mammal such as increasing phagocytosis, for example, prolonging the life of phagocytes,
inter alia, keratinocytes, neutrophils. As such the disclosed compounds provide methods for treating
diseases that relate to the body's immune response.
SUMMARY
[0003] The disclosed compounds stabilize HIF-1α and HIF-2α, as well as other factors that
are present in a compromised immune system or which are depleted or over taxed by
the presence of a disease state and the manifestations of the disease state,
inter alia, sepsis. The disclosed compounds can be used to treat cancer and can be co-administered
with other cancer therapy drugs. In addition, the disclosed compounds can be used
to boost the immune response by a mammal when co-administered with a vaccine, for
example, flu vaccines, malarial vaccines, yellow fever vaccines, cancer vaccines,
and the like.
BRIEF DESCRIPTION OF THE FIGURES
[0004]
Figure 1 depicts the normal metabolic pathway of HIF-1α during normoxia.
Figure 2 depicts the enhancement of neutrophil killing of S. aureus (Newman strain) with 50 µM and 200 µM of a compound disclosed in Table VIII versus
control (DMSO) at 60 and 90 minutes.
Figure 3 depicts the enhancement of human monocyte cell line (U937) against S. aureus (Newman strain) by 10 µM a compound disclosed in Table VIII versus untreated samples.
Figure 4 depicts the average percent surviving bacteria in treated vs. untreated U937 cells
after infection with S. aureus (Newman strain) after 1 hour pre-treatment (black) or 2 hour (hatched) pre-treatment
with 10 µM of a compound disclosed in Table VIII.
Figure 5 depicts the average percent surviving bacteria in treated vs. untreated U937 cells
after infection with two strains of S. aureus, Newman (black) or methicillin resistant S. aureus (MRSA) (hatched), after 1 hour pre-treatment with 10 µM of a compound disclosed in
Table VIII.
Figure 6 depicts the average percent surviving bacteria in treated vs. untreated U397 cells
after infection with two strains of S. aureus, Newman (black) or MRSA (hatched) and treatment with 10 µM of a compound disclosed
in Table VIII.
Figure 7 depicts the average percent surviving bacteria in treated vs. untreated U937 cells
after infection with two strains of S. aureus, Newman (hatched bars) or MRSA (black bars), following treatment with 100 mM mimosine
(A), 10 µM of a compound disclosed in Table VIII (B), or 2mg/mL of vancomycin (C)
at 2 hours post-infection.
Figure 8 depicts the average percent surviving bacteria in treated vs. untreated U937 cells
after infection with S. aureus (Newman) following no pre-treatment, 1 hour pre-treatment, or 2 hour pre-treatment
with 10 µM of a compound disclosed in Table VIII.
Figure 9 depicts the average percent surviving bacteria in treated vs. untreated HaCaT cells
infected with two strains of S. aureus, Newman (hatched bars) or MRSA (black bars) and pre-treated for 1 hour with either
DMSO (control), 800 µM mimosine, 10 µM a compound disclosed in Table VIII or 1 µg/mL
vancomycin. Data shown is 2 hours post-treatment.
Figure 10 depicts the average percent surviving bacteria in treated vs. untreated HaCaT cells
infected with two strains of S. aureus, Newman (hatched bars) or MRSA (black bars), following pre-treatment with 10 µM a
compound disclosed in Table VIII.
Figure 11 depicts the up regulation of phosphoglycerate kinase (PGK) expression in wild type
murine embryonic fibroblasts as a result of treatment with a compound disclosed in
Table VIII at dosages of 1 µM (E), 10 µM (F), and 50 µM (G) vs. wild type control
(H) and the lack of up regulation of PGK expression in HIF-1 knock out cells as a
result of treatment with a compound disclosed in Table VIII at dosages of 1 µM (A),
10 µM (B), and 50 µM (C) and HIF-1 knock out control (D). Both cell types were treated
for 7 hours.
Figure 12 depicts the up regulation of phosphoglycerate kinase (PGK) expression in wild type
murine embryonic fibroblasts as a result of treatment with compound 1-(3-Chlorobenzyl)-3-hydroxypyridin-2(1H)-one
at dosages of 1 µM (E), 10 µM (F), vs. wild type control (G) and the lack of up regulation
of PGK expression in HIF-1 knock out cells as a result of treatment with a compound
disclosed in Table VIII at dosages of 1 µM (A), 10 µM (B), and 50 µM (C) and HIF-1
knock out control (D).
Figure 13 depicts the up regulation of phosphoglycerate kinase (PGK) expression in wild type
murine embryonic fibroblasts as a result of treatment with compound a compound disclosed
in Table VIII at dosages of 1 µM (E), 10 µM (F), and 50 µM (G) vs. wild type control
(H) and the lack of up regulation of PGK expression in HIF-1 knock out cells as a
result of treatment with compound a compound disclosed in Table VIII at dosages of
1 µM (A), 10 µM (B), and 50 µM (C) and HIF-1 knock out control (D).
Figure 14 depicts the up regulation of vascular endothelia growth factor (VEGF) expression
in wild type murine embryonic fibroblasts as a result of treatment with compound a
compound disclosed in Table VIII at dosages of 1 µM (E), 10 µM (F), and 50 µM (G)
vs. control (H) and the lack of up regulation of VEGF expression in HIF-1 knock out
cells treated with a compound disclosed in Table VIII at dosages of 1 µM (A), 10 µM
(B), and 50 µM (C) and HIF-1 knock out control (D). Both cell types were treated for
7 hours.
Figure 15 depicts the results of Example 11 wherein 3 groups of animals are treated with Staphylococcus aureus antibiotic sensitive Newman strain. The data show the significant reduction in the
size of skin lesions (wounds) for animals in Group 1 (solid circles (●)) treated with
10µM of a compound disclosed in Table VIII versus animal given a bolus of DMSO (solid
squares (■)). Figure 15, depicts mice infected with Newman strain of S. aureus followed
by treatment with 10 µM of a compound disclosed in Table VIII or DMSO (control) at
2 hours post-infection. The data show the statistically significant reduction in the
size of skin lesions (wounds) for animals treated with compound a compound disclosed
in Table VIII (solid circles (●)) or DMSO (solid squares (■)).
Figure 16 also depicts the results of Example 11 showing the reduction in the size of skin
lesions (wounds) for animals in Group 1 (solid circles (●)) treated with 10µM a compound
disclosed in Table VIII versus animals that are untreated (solid triangles (▲)). Figure 16 depicts mice infected with Newman strain of S. aureus followed by treatment with
10 µM of a compound disclosed in Table VIII or no treatment at 2 hours post-infection.
The data show the reduction in the size of skin lesions (wounds) for animals treated
with compound a compound disclosed in Table VIII (solid circles (●)) or untreated
(solid triangles (A)).
Figure 17 is a plot histogram that depicts the results of Example 12 wherein 3 groups of animals
are treated with Staphylococcus aureus antibiotic sensitive Newman strain [ATCC #25904]. The data show the results for the
untreated group plotted under (A), the results for the group treated with DMSO plotted
under (B) and results for the group treated with 10µM of a compound disclosed in Table
VIII plotted under (C).
Figure 18 also depicts the results of Example 12 wherein the number of colony forming units
in the kidney are plotted for the various groups: the untreated group is plotted under
(A), the group treated with DMSO is plotted under (B) and the group treated with 10µM
of a compound disclosed in Table VIII is plotted under (C).
Figure 19 depicts the results of Example 13 wherein 2 groups of animals are treated with Streptococcus pyogenes NZ131 [M49 strain]. The data show the reduction in the size of skin lesions (wounds)
for animals in Group 1 (solid triangles (A)) treated with 0.5 mg/kg of a compound
disclosed in Table VIII versus animal treated with vehicle control (cyclodextran)
(solid circles (●)).
Figure 20 is a plot histogram that also depicts the results of Example 12 wherein the number
of colony forming units for the observed skin lesions on animals treated with vehicle
control (cyclodextran) are plotted under (A) and results for the group treated with
0.5 mg/kg of a compound disclosed in Table VIII are plotted under (B).
DETAILED DISCLOSURE
[0005] In this specification and in the claims that follow, reference will be made to a
number of terms that shall be defined to have the following meanings:
[0006] Throughout this specification, unless the context requires otherwise, the word "comprise,"
or variations such as "comprises" or "comprising," will be understood to imply the
inclusion of a stated integer or step or group of integers or steps but not the exclusion
of any other integer or step or group of integers or steps.
[0007] It must be noted that, as used in the specification and the appended claims, the
singular forms "a," "an" and "the" include plural referents unless the context clearly
dictates otherwise. Thus, for example, reference to "a carrier" includes mixtures
of two or more such carriers, and the like.
[0008] "Optional" or "optionally" means that the subsequently described event or circumstance
can or cannot occur, and that the description includes instances where the event or
circumstance occurs and instances where it does not.
[0009] By "pharmaceutically acceptable" is meant a material that is not biologically or
otherwise undesirable, i.e., the material can be administered to an individual along
with the relevant active compound without causing clinically unacceptable biological
effects or interacting in a deleterious manner with any of the other components of
the pharmaceutical composition in which it is contained.Ranges may be expressed herein
as from "about" one particular value, and/or to "about" another particular value.
When such a range is expressed, another aspect includes from the one particular value
and/or to the other particular value. Similarly, when values are expressed as approximations,
by use of the antecedent "about," it will be understood that the particular value
forms another aspect. It will be further understood that the endpoints of each of
the ranges are significant both in relation to the other endpoint, and independently
of the other endpoint.
[0010] A weight percent of a component, unless specifically stated to the contrary, is based
on the total weight of the formulation or composition in which the component is included.
[0011] By "effective amount" as used herein means "an amount of one or more of the disclosed
HIF-1α prolyl hydroxylase inhibitors, effective at dosages and for periods of time
necessary to achieve the desired or therapeutic result." An effective amount may vary
according to factors known in the art, such as the disease state, age, sex, and weight
of the human or animal being treated. Although particular dosage regimes may be described
in examples herein, a person skilled in the art would appreciated that the dosage
regime may be altered to provide optimum therapeutic response. For example, several
divided doses may be administered daily or the dose may be proportionally reduced
as indicated by the exigencies of the therapeutic situation. In addition, the compositions
of this disclosure can be administered as frequently as necessary to achieve a therapeutic
amount.
[0012] "Admixture" or "blend" is generally used herein means a physical combination of two
or more different components
[0013] "Excipient" is used herein to include any other compound that may be contained in
or combined with one or more of the disclosed inhibitors that is not a therapeutically
or biologically active compound. As such, an excipient should be pharmaceutically
or biologically acceptable or relevant (for example, an excipient should generally
be non-toxic to the subject). "Excipient" includes a single such compound and is also
intended to include a plurality of excipients.
[0014] As used herein, by a "subject" is meant an individual. Thus, the "subject" can include
domesticated animals (
e.g., cats, dogs, etc.), livestock (
e.g., cattle, horses, pigs, sheep, goats, etc.), laboratory animals (
e.g., mouse, rabbit, rat, guinea pig, etc.), and birds. "Subject" can also include a mammal,
such as a primate or a human.
[0015] By "prevent" or other forms of the word, such as "preventing" or "prevention," is
meant to stop a particular event or characteristic, to stabilize or delay the development
or progression of a particular event or characteristic, or to minimize the chances
that a particular event or characteristic will occur. Prevent does not require comparison
to a control as it is typically more absolute than, for example, reduce. As used herein,
something could be reduced but not prevented, but something that is reduced could
also be prevented. Likewise, something could be prevented but not reduced, but something
that is prevented could also be reduced. It is understood that where reduce or prevent
are used, unless specifically indicated otherwise, the use of the other word is also
expressly disclosed.
[0016] By "reduce" or other forms of the word, such as "reducing" or "reduction," is meant
lowering of an event or characteristic (
e.g., vascular leakage). It is understood that this is typically in relation to some standard
or expected value, in other words it is relative, but that it is not always necessary
for the standard or relative value to be referred to.
[0017] The term "Treat" or other forms of the word such as "treated" or "treatment" is used
herein to mean that administration of a compound of the present invention mitigates
a disease or a disorder in a host and/or reduces, inhibits, or eliminates a particular
characteristic or event associated with a disorder (e.g., infection caused by a microorganism).
Thus, the term "treatment" includes, preventing a disorder from occurring in a host,
particularly when the host is predisposed to acquiring the disease, but has not yet
been diagnosed with the disease; inhibiting the disorder; and/or alleviating or reversing
the disorder. Insofar as the methods of the present invention are directed to preventing
disorders, it is understood that the term "prevent" does not require that the disease
state be completely thwarted. Rather, as used herein, the term preventing refers to
the ability of the skilled artisan to identify a population that is susceptible to
disorders, such that administration of the compounds of the present invention may
occur prior to onset of a disease. The term does not imply that the disease state
be completely avoided.
[0018] Ranges can be expressed herein as from "about" one particular value, and/or to "about"
another particular value. When such a range is expressed, another aspect includes
from the one particular value and/or to the other particular value. Similarly, when
values are expressed as approximations, by use of the antecedent "about," it will
be understood that the particular value forms another aspect. It will be further understood
that the endpoints of each of the ranges are significant both in relation to the other
endpoint, and independently of the other endpoint. It is also understood that there
are a number of values disclosed herein, and that each value is also herein disclosed
as "about" that particular value in addition to the value itself. For example, if
the value "10" is disclosed, then "about 10" is also disclosed. It is also understood
that when a value is disclosed, then "less than or equal to" the value, "greater than
or equal to the value," and possible ranges between values are also disclosed, as
appropriately understood by the skilled artisan. For example, if the value "10" is
disclosed, then "less than or equal to 10" as well as "greater than or equal to 10"
is also disclosed. It is also understood that throughout the application data are
provided in a number of different formats and that this data represent endpoints and
starting points and ranges for any combination of the data points. For example, if
a particular data point "10" and a particular data point "15" are disclosed, it is
understood that greater than, greater than or equal to, less than, less than or equal
to, and equal to 10 and 15 are considered disclosed as well as between 10 and 15.
It is also understood that each unit between two particular units are also disclosed.
For example, if 10 and 15 are disclosed, then 11, 12, 13, and 14 are also disclosed.By
"antimicrobial" is meant the ability to treat or control (
e.g., reduce, prevent, inhibit, break-down, or eliminate) microorganism growth or survival
at any concentration. Similarly, the terms "antibacterial," "antiviral," and "antifungal"
respectively mean the ability to treat or control (
e.g., reduce, prevent, inhibit, break-down, or eliminate) bacterial, viral, and fungal
growth or survival at any concentration.
[0019] The term "anion" is a type of ion and is included within the meaning of the term
"ion". An "anion" is any molecule, portion of a molecule (
e.g., zwitterion), cluster of molecules, molecular complex, moiety, or atom that contains
a net negative charge or that can be made to contain a net negative charge. The term
"anion precursor" is used herein to specifically refer to a molecule that can be converted
to an anion via a chemical reaction (
e.g., deprotonation).
[0020] The term "cation" is a type of ion and is included within the meaning of the term
"ion". A "cation" is any molecule, portion of a molecule (
e.g., zwitterion), cluster of molecules, molecular complex, moiety, or atom, that contains
a net positive charge or that can be made to contain a net positive charge. The term
"cation precursor" is used herein to specifically refer to a molecule that can be
converted to a cation via a chemical reaction (
e.g., protonation or alkylation).
[0021] "Chemotherapeutic agent" is used herein to include any other pharmaceutically active
compound that can be used in conjunction with the disclosed HIF-1α prolyl hydroxylase
inhibitors, for example, cytotoxic drugs such as 6-hydroxymethylacylfulvene, cyclophosphamide,
dacarbazine, carmustine, doxorubicin, and methotrexate. Other chemotherapeutic agents
also include anti-inflammatory drugs,
i.e., non-steroidal anti-inflammatory compounds such as aspirin.
[0022] Unless stated to the contrary, a formula with chemical bonds shown only as solid
lines and not as wedges or dashed lines contemplates each possible isomer,
e.g., each enantiomer, diastereomer, and meso compound, and a mixture of isomers, such
as a racemic or scalemic mixture.
[0023] The transcription factor Hypoxia-Inducible Factor 1 (HIF-1) is one of the key regulators
of oxygen homeostasis. It regulates the physiological responses to low oxygen levels
(hypoxia) and the pathophysiology of heart attack, cancer, stroke and chronic lung
disease. HIF-1 is a heterodimeric protein that consists of two subunits, HIF-1α and
HIF-1β. Whereas HIF-1β is constitutively expressed, the expression of HIF-1α is induced
by oxygen concentrations below 6%. HIF-1 heterodimers bind to the hypoxia response
element (HRE), a 5-RCGTG-3 consensus sequence. Several dozen HIF-1-regulated genes
have been identified so far, including genes coding for proteins involved in angiogenesis,
energy metabolism, erythropoiesis, cell proliferation and viability, vascular remodeling
and vasomotor responses. Therefore, modulation of HIF activation in cells is critical
to preventing, controlling, curing, or otherwise affecting a wide array of diseases,
disease states, and conditions.
[0024] Hypoxia-inducible transcription factor 1-alpha (HIF-1α) plays a central role in cellular
adaptation to reduced oxygen availability. Under hypoxic stress, activated HIF-1α
strives for oxygen homeostasis by not only maintaining intracellular energy production
via the induction of angiogenesis and glycolysis, but also limiting energy consumption
by virtue of the inhibition of cell proliferation and DNA repair. In general, HIF-1α
activates its target genes,
inter alia, EPO, VEGF, and
PGK1 through binding to the hypoxia-responsive element in the gene promoter (
Wang, G.L. et al., J Biol Chem (1993); 268: 21513-21518).
[0025] HIF-1α under normal healthy conditions wherein the cells have a sufficient supply
of oxygen is readily converted to a degraded form by one of several 4-prolyl hydroxylase
enzymes,
inter alia, EGLN1 (herein referred to as HIFPH2). As stated above, when cells undergo hypoxia,
this enzymatic transformation is slow or entirely stopped and HIF-1α begins to build
up in the cell. When this build up of HIF-1α occurs, this protein combines with HIF-1β
to form the active transcription factor complex HIF-1. This transcription factor then
activates several biological pathways which are present as a response to and a means
for alleviating the body's state of hypoxia. These responses include,
inter alia, angiogenic, erythropoietic (EPO), glucose metabolism (PGK), matrix alteration, and
enhanced capacity of phagocytes to respond to pathogens.
[0026] Figure 1 summaries the metabolism of HIF-1α during normal healthy conditions. The HIF α-subunits
are unstable under normoxic conditions; cells continually synthesize and degrade these
proteins. The short half-life of HIF-1α is the byproduct of a family of O
2-and iron-dependent prolyl hydroxylases (pH1-3), whose action directs HIF α-subunits
for degradation by the ubiquitin-proteasome pathway in a process dependent upon interaction
with von Hippel-Lindau tumor-suppressor protein (vHL). In
Figure 1, PDH's represents the prolyl hydroxylases that act in the presence of an asparaginyl
hydroxylase to hydroxylate prolines 402 and 564, as well as asparagines 804. From
this point, because the hydroxylated HIF-1α is also prevented from association with
p300-CPB because of other factors, ubiquitin ligase begins to metabolize the hydroxylated
HIF-1α via the vHL pathway.
[0027] In patients where there is a need for stimulating this response, for example, in
patients in need of increased tissue oxygen due to peripheral vascular disease (PVD),
inhibiting the HIF1 enzymes, for example, Egl nine homolog 1 (HIFPH2), will stimulate
the body's own angiogenic response without the consequences of oxygen deficiency.
In addition, in diseases of ischemia,
inter alia, CAD and anemia, stimulation of angiogenic, erythropoietic, and metabolism adaption
can provide therapeutic benefits. Up regulation of HIF-1α also provides a method for
enhancing immunity, for example, by increasing the capacity of phagocytes.
[0028] There is therefore a long felt need for methods for controlling the activity of HIF-1α
which can be effectively accomplished by compounds that inhibit the 4-prolyl hydroxylase
enzymes that degrade HIF-1α. This inhibition of 4-prolyl hydroxylases enzymes,
inter alia, HIFPH2 (also referred to herein as EGLN1 or PHD2) and HIFPH3 (also referred to herein
as EGLN3 of PHD-3) thereby provide a method for increasing the concentration of HIF-1α
in cells and thus providing methods for treating a variety of diseases or disease
states.
[0029] Disclosed herein are methods for treating one or more diseases, conditions, syndromes,
and the like that are affected by the level of hypoxia-inducible transcription factors.
Regulation of these factors both during hypoxia and normoxia can provide methods for
re-balancing or regulating one or more biological pathways associated with abnormal
conditions,
inter alia, invasion of the body by pathogens,
inter alia, bacteria, fungi, viruses, and parasites, abnormal cellular regulation,
i.e., cancer ischemia, and the side effects caused by vaccination.
Targeting HIF1 Stabilization in Cells
[0030] HIF-1α is targeted for destruction via prolyl hydroxylation, an oxygen-dependent
modification that signals for recognition by the E3 ubiquitin ligase complex containing
the von Hippel-Lindau tumor suppressor (VHL). Three prolyl hydroxylases formerly referred
to in the literature as EGLN1, EGLN2, and EGLN3 (also know as , have been identified
in mammals, among which, EGLN1 (also known as HIFPH2 or PHD2), and EGLN3 (also known
as HIFPH3 or PHD3), are hypoxia-inducible at their mRNA levels in a HIF-1α dependent
manner. HIF-1α levels are controlled by these prolyl-4-hydroxylases by hydroxylating
the HIF-1α proline residues Pro-402 and Pro-564 in humans (
Ivan, M. et al., (2001) "HIFα targeted for VHL-mediated destruction by proline hydroxylation:
implications for O2 sensing." Science 292, 464-468;
Jaakkola, P. et al., (2001) "Targeting of HIF-1α to the von Hippel-Lindau ubiquitylation
complex by O2-regulated prolyl hydroxylation." Science 292, 468-472; and
Masson, N. et al., (2001) "Independent function of two destruction domains in hypoxia-inducible
factor-α chains activated by prolyl hydroxylation." EMBO J. 20, 5197-5206). Under hypoxia conditions, EGLN1 and EGLN3 activity is suppressed.
[0031] Stimulated by a build up of the cellular concentration of HIF-1α is the production
of Phosphoglycerate Kinase (PGK) and Vascular Endothelial Growth Factor (VEGF). It
has been shown that stimulation of VEGF induces the formation of functional neo-vessels
in the mouse cornea and enhanced blood flow in a dog model of coronary artery disease.
The HIF-1α prolyl hydroxylase inhibitors of the present disclosure provide enhancement
in the expression of multiple hypoxia inducible genes including VEGF, GAPDH and erythropoietin
(EPO). Additionally, the HIF-1α prolyl hydroxylase inhibitors of the present disclosure
provide enhanced the accumulation of HIF1-α in the cytoplasm and nucleus. Transgenic
mice expressing a constitutively active HIF-1α in the skin have increased dermal vascularity
and had a 13-fold increase in VEGF levels
Wounds
[0032] Chronic, non-healing wounds are a major cause of prolonged morbidity in the aged
human population. This is especially the case in bedridden or diabetic patients who
develop severe, non-healing skin ulcers. In many of these cases, the delay in healing
is a result of inadequate blood supply either as a result of continuous pressure or
of vascular blockage. Poor capillary circulation due to small artery atherosclerosis
or venous stasis contributes to the failure to repair damaged tissue. Such tissues
are often infected with microorganisms that proliferate unchallenged by the innate
defense systems of the body which require well vascularized tissue to effectively
eliminate pathogenic organisms. As a result, most therapeutic intervention centers
on restoring blood flow to ischemic tissues thereby allowing nutrients and immunological
factors access to the site of the wound.
[0033] The present disclosure relates to methods for treating wounds and promoting wound
healing in a subject comprising, administering to a subject in need of treatment an
effective amount of one or more of the disclosed compounds.
[0034] The present disclosure relates to the use of one or more of the disclosed compounds
for use in making a medicament for treating wounds and promoting wound healing.
Antimicrobial
[0035] The hypoxia-responsive transcription factor HIF-1α is essential for regulation of
inflammation
in vivo. As such, it has been discovered (
Peyssonnaux C. et al., "HIF-1α expression regulates the bactericidal capacity of phagocytes"
J. Clinical Investigation 115(7), pp 1808- 1815 (2005)) that bacterial infection induces HIF-1α expression in myeloid cells even under
normoxic conditions, and that HIF-1α regulates the generation of critical molecular
effectors of immune defense including granule proteases, antimicrobial peptides, nitric
oxide, and TNF-α. Bacterial infection induces a subset of HIF-1α target genes specifically
related to microbial killing, thereby demonstrating that HIF-1α has an essential function
in innate immunity distinct from hypoxic response. Therefore, HIF-1α function is critical
for myeloid cell bactericidal activity and the ability of the host to limit systemic
spread of infection from an initial tissue focus. Increased activity of the HIF-1α
pathway through vHL deletion supports myeloid cell production of defense factors and
improves bactericidal capacity. The disclosed compounds induce HIF-1α activity and
can also boost bacterial killing and NO production in a HIF-1α-specific fashion. These
discoveries provide methods for enhancing innate immune responses to microbial, for
example, bacterial, infection.
[0036] Without wishing to be limited by theory, the disclosed compounds can increase the
stabilization of HIF-1 protein by acting directly or indirectly on one or more cellular
processes which act to destabilize or to metabolize cellular components that stabilize
the presence of HIF-1 protein, protect it from inhibition, or to increase the activity
of the protein. Alternatively, the disclosed compounds can increase the activity of
the HIF-1 protein by inhibiting or otherwise blocking the activity of compounds that
inhibit the activity of the HIF-1 protein. As such, disclosed herein is a method for
improving the treatment of microbial infections by administering a substance that
increases the activity or level of at least one HIF-1 protein in a subject suffering
from the microbial infection or at increased risk of microbial infection.
[0037] In one aspect, disclosed herein are methods for modulating the activity of at least
one HIF-1 protein. As such, the disclosed methods comprise contacting at least one
HIF-1 protein or HIF-1 interacting protein with one or more of the disclosed compounds
that modulate the activity of the HIF-1 protein, or causing contact between the protein
and substance. In the embodiment, the contacting is accomplished
in vitro. In another embodiment, the contacting is accomplished
in vivo. In a further embodiment, the contacting is accomplished
ex vivo.
[0038] In another aspect, disclosed herein is a method of treating a subject infected or
at risk of infection by a microbial agent comprising administering to a subject a
therapeutically effective amount of one or more of the disclosed compounds. In one
embodiment, the compound increases the amount or activity of HIF-1. In another embodiment,
the microbial agent is a pathogen. Iterations of this embodiment related to pathogens
includes, bacteria, fungi, protozoa, viruses, yeasts, and the like. A yet further
iteration of this aspect relates to a method for treating a subject infected by or
at risk of infection by a microbial agent comprising, increasing the microbial pathogen-killing
activity of the subject's immune cells.
[0039] One method for increasing the stabilization of HIF-1 is to inhibit the activity of
4-prolyl hydroxylase enzymes that begin the cellular break down of HIF-1α thereby
preventing HIF-1α from combining with HIF-1β to form HIF-1. As such, disclosed herein
are methods for increasing the cellular response to disease states such as infection,
i.e., presence of a pathogen such as a bacterium, a virus, a parasite, a yeast, a
fungus, and the like by increasing phagocytosis. Also disclosed herein are methods
for treating cancer by increasing the cellular immune response, for example, by stabilizing
HIF-1, thereby increasing the ability of the body to reduce tumor size. Further disclosed
herein are methods for treating diseases wherein an immune response can be stimulated
by vaccination.
[0040] The following chemical hierarchy is used throughout the specification to describe
and enable the scope of the present disclosure and to particularly point out and distinctly
claim the units which comprise the compounds of the present disclosure, however, unless
otherwise specifically defined, the terms used herein are the same as those of the
artisan of ordinary skill. The term "hydrocarbyl" stands for any carbon atom-based
unit (organic molecule), said units optionally containing one or more organic functional
group, including inorganic atom comprising salts,
inter alia, carboxylate salts, quaternary ammonium salts. Within the broad meaning of the term
"hydrocarbyl" are the classes "acyclic hydrocarbyl" and "cyclic hydrocarbyl" which
terms are used to divide hydrocarbyl units into cyclic and non-cyclic classes.
[0041] As it relates to the following definitions, "cyclic hydrocarbyl" units can comprise
only carbon atoms in the ring (carbocyclic and aryl rings) or can comprise one or
more heteroatoms in the ring (heterocyclic and heteroaryl). For "carbocyclic" rings
the lowest number of carbon atoms in a ring are 3 carbon atoms; cyclopropyl. For "aryl"
rings the lowest number of carbon atoms in a ring are 6 carbon atoms; phenyl. For
"heterocyclic" rings the lowest number of carbon atoms in a ring is 1 carbon atom;
diazirinyl. Ethylene oxide comprises 2 carbon atoms and is a C
2 heterocycle. For "heteroaryl" rings the lowest number of carbon atoms in a ring is
1 carbon atom; 1,2,3,4-tetrazolyl. The following is a non-limiting description of
the terms "acyclic hydrocarbyl" and "cyclic hydrocarbyl" as used herein.
- A. Substituted and unsubstituted acyclic hydrocarbyl:
For the purposes of the present disclosure the term "substituted and unsubstituted
acyclic hydrocarbyl" encompasses 3 categories of units:
- 1) linear or branched alkyl, non-limiting examples of which include, methyl (C1), ethyl (C2), n-propyl (C3), iso-propyl (C3), n-butyl (C4), sec-butyl (C4), iso-butyl (C4), tert-butyl (C4), and the like; substituted linear or branched alkyl, non-limiting examples of which
includes, hydroxymethyl (C1), chloromethyl (C1), trifluoromethyl (C1), aminomethyl (C1), 1-chloroethyl (C2), 2-hydroxyethyl (C2), 1,2-difluoroethyl (C2), 3-carboxypropyl (C3), and the like.
- 2) linear or branched alkenyl, non-limiting examples of which include, ethenyl (C2), 3-propenyl (C3), 1-propenyl (also 2-methylethenyl) (C3), isopropenyl (also 2-methylethen-2-yl) (C3), buten-4-yl (C4), and the like; substituted linear or branched alkenyl, non-limiting examples of
which include, 2-chloroethenyl (also 2-chlorovinyl) (C2), 4-hydroxybuten-1-yl (C4), 7-hydroxy-7-methyloct-4-en-2-yl (C9), 7-hydroxy-7-methyloct-3,5-dien-2-yl (C9), and the like.
- 3) linear or branched alkynyl, non-limiting examples of which include, ethynyl (C2), prop-2-ynyl (also propargyl) (C3), propyn-1-yl (C3), and 2-methyl-hex-4-yn-1-yl (C7); substituted linear or branched alkynyl, non-limiting examples of which include,
5-hydroxy-5-methylhex-3-ynyl (C7), 6-hydroxy-6-methylhept-3-yn-2-yl (C8), 5-hydroxy-5-ethylhept-3-ynyl (C9), and the like.
B. Substituted and unsubstituted cyclic hydrocarbyl:
For the purposes of the present disclosure the term "substituted and unsubstituted
cyclic hydrocarbyl" encompasses 5 categories of units:
- 1) The term "carbocyclic" is defined herein as "encompassing rings comprising from
3 to 20 carbon atoms, wherein the atoms which comprise said rings are limited to carbon
atoms, and further each ring can be independently substituted with one or more moieties
capable of replacing one or more hydrogen atoms." The following are non-limiting examples
of "substituted and unsubstituted carbocyclic rings" which encompass the following
categories of units:
- i) carbocyclic rings having a single substituted or unsubstituted hydrocarbon ring,
non-limiting examples of which include, cyclopropyl (C3), 2-methyl-cyclopropyl (C3), cyclopropenyl (C3), cyclobutyl (C4), 2,3-dihydroxycyclobutyl (C4), cyclobutenyl (C4), cyclopentyl (C5), cyclopentenyl (C5), cyclopentadienyl (C5), cyclohexyl (C6), cyclohexenyl (C6), cycloheptyl (C7), cyclooctanyl (C8), 2,5-dimethylcyclopentyl (C5), 3,5-dichlorocyclohexyl (C6), 4-hydroxycyclohexyl (C6), and 3,3,5-trimethylcyclohex-1-yl (C6).
- ii) carbocyclic rings having two or more substituted or unsubstituted fused hydrocarbon
rings, non-limiting examples of which include, octahydropentalenyl (C8), octahydro-1H-indenyl (C9), 3a,4,5,6,7,7a-hexahydro-3H-inden-4-yl (C9), decalinyl (C10), decahydroazulenyl (C10).
- iii) carbocyclic rings which are substituted or unsubstituted bicyclic hydrocarbon
rings, non-limiting examples of which include, bicyclo-[2.1.1]hexanyl, bicyclo[2.2.1]heptanyl,
bicyclo[3.1.1]heptanyl, 1,3-dimethyl[2.2.1]heptan-2-yl, bicyclo[2.2.2]octanyl, and
bicyclo[3.3.3]undecanyl.
- 2) The term "aryl" is defined herein as "units encompassing at least one phenyl or
naphthyl ring and wherein there are no heteroaryl or heterocyclic rings fused to the
phenyl or naphthyl ring and further each ring can be independently substituted with
one or more moieties capable of replacing one or more hydrogen atoms." The following
are non-limiting examples of "substituted and unsubstituted aryl rings" which encompass
the following categories of units:
- i) C6 or C10 substituted or unsubstituted aryl rings; phenyl and naphthyl rings whether substituted
or unsubstituted, non-limiting examples of which include, phenyl (C6), naphthylen-1-yl (C10), naphthylen-2-yl (C10), 4-fluorophenyl (C6), 2-hydroxyphenyl (C6), 3-methylphenyl (C6), 2-amino-4-fluorophenyl (C6), 2-(N,N-diethylamino)phenyl (C6), 2-cyanophenyl (C6), 2,6-di-tert-butylphenyl (C6), 3-methoxyphenyl (C6), 8-hydroxynaphthylen-2-yl (C10), 4,5-dimethoxynaphthylen-1-yl (C10), and 6-cyano-naphthylen-1-yl (C10).
- ii) C6 or C10 aryl rings fused with 1 or 2 saturated rings non-limiting examples of which include,
bicyclo[4.2.0]octa-1,3,5-trienyl (C8), and indanyl (C9).
- 3) The terms "heterocyclic" and/or "heterocycle" are defined herein as "units comprising
one or more rings having from 3 to 20 atoms wherein at least one atom in at least
one ring is a heteroatom chosen from nitrogen (N), oxygen (O), or sulfur (S), or mixtures
of N, O, and S, and wherein further the ring which comprises the heteroatom is also
not an aromatic ring." The following are non-limiting examples of "substituted and
unsubstituted heterocyclic rings" which encompass the following categories of units:
- i) heterocyclic units having a single ring containing one or more heteroatoms, non-limiting
examples of which include, diazirinyl (C1), aziridinyl (C2), urazolyl (C2), azetidinyl (C3), pyrazolidinyl (C3), imidazolidinyl (C3), oxazolidinyl (C3), isoxazolinyl (C3), thiazolidinyl (C3), isothiazolinyl (C3), oxathiazolidinonyl (C3), oxazolidinonyl (C3), hydantoinyl (C3), tetrahydrofuranyl (C4), pyrrolidinyl (C4), morpholinyl (C4), piperazinyl (C4), piperidinyl (C4), dihydropyranyl (C5), tetrahydropyranyl (C5),piperidin-2-onyl (valerolactam) (C5), 2,3,4,5-tetrahydro-1H-azepinyl (C6), 2,3-dihydro-1H-indole (C8), and 1,2,3,4-tetrahydro-quinoline (C9).
- ii) heterocyclic units having 2 or more rings one of which is a heterocyclic ring,
non-limiting examples of which include hexahydro-1H-pyrrolizinyl (C7), 3a,4,5,6,7,7a-hexahydro-1H-benzo[d]imidazolyl (C7), 3a,4,5,6,7,7a-hexahydro-1H-indolyl (C8), 1,2,3,4-tetrahydroquinolinyl (C9), and decahydro-1H-cycloocta[b]pyrrolyl (C10).
- 4) The term "heteroaryl" is defined herein as "encompassing one or more rings comprising
from 5 to 20 atoms wherein at least one atom in at least one ring is a heteroatom
chosen from nitrogen (N), oxygen (O), or sulfur (S), or mixtures ofN, O, and S, and
wherein further at least one of the rings which comprises a heteroatom is an aromatic
ring." The following are non-limiting examples of "substituted and unsubstituted heterocyclic
rings" which encompass the following categories of units:
- i) heteroaryl rings containing a single ring, non-limiting examples of which include,
1,2,3,4-tetrazolyl (C1), [1,2,3]triazolyl (C2), [1,2,4]triazolyl (C2), triazinyl (C3), thiazolyl (C3), 1H-imidazolyl (C3), oxazolyl (C3), isoxazolyl (C3), isothiazolyl (C3), furanyl (C4), thiopheneyl (C4), pyrimidinyl (C4), 2-phenylpyrimidinyl (C4), pyridinyl (C5), 3-methylpyridinyl (C5), and 4-dimethylaminopyridinyl (C5).
- ii) heteroaryl rings containing 2 or more fused rings one of which is a heteroaryl
ring, non-limiting examples of which include: 7H-purinyl (C5), 9H-purinyl (C5), 6-amino-9H-purinyl (C5), 5H-pyrrolo[3,2-d]pyrimidinyl (C6), 7H-pyrrolo[2,3-d]pyrimidinyl (C6), pyrido[2,3-d]pyrimidinyl (C7), 2-phenylbenzo[d]thiazolyl (C7), 1H-indolyl (C8), 4,5,6,7-tetrahydro-1-H-indolyl (C8), quinoxalinyl (C8), 5-methylquinoxalinyl (C8), quinazolinyl (C8), quinolinyl (C9), 8-hydroxy-quinolinyl (C9), and isoquinolinyl (C9).
- 5) C1-C6 tethered cyclic hydrocarbyl units (whether carbocyclic units, C6 or C10 aryl units, heterocyclic units, or heteroaryl units) which connected to another moiety,
unit, or core of the molecule by way of a C1-C6 alkylene unit. Non-limiting examples of tethered cyclic hydrocarbyl units include
benzyl C1-(C6) having the formula:
wherein R
a is optionally one or more independently chosen substitutions for hydrogen. Further
examples include other aryl units,
inter alia, (2-hydroxyphenyl)hexyl C
6-(C
6); naphthalen-2-ylmethyl C
1-(C
10), 4-fluorobenzyl C
1-(C
6), 2-(3-hydroxy-phenyl)ethyl C
2-(C
6), as well as substituted and unsubstituted C
3-C
10 alkylenecarbocyclic units, for example, cyclopropylmethyl C
1-(C
3), cyclopentylethyl C
2-(C
5), cyclohexylmethyl C
1-(C
6);. Included within this category are substituted and unsubstituted C
1-C
10 alkylene-heteroaryl units, for example a 2-picolyl C
1-(C
6) unit having the formula:
wherein R
a is the same as defined above. In addition, C
1-C
12 tethered cyclic hydrocarbyl units include C
1-C
10 alkyleneheterocyclic units and alkylene-heteroaryl units, non-limiting examples of
which include, aziridinylmethyl C
1-(C
2) and oxazol-2-ylmethyl C
1-(C
3).
[0042] For the purposes of the present disclosure carbocyclic rings are from C
3 to C
20; aryl rings are C
6 or C
10; heterocyclic rings are from C
1 to C
9; and heteroaryl rings are from C
1 to C
9.
[0043] For the purposes of the present disclosure, and to provide consistency in defining
the present disclosure, fused ring units, as well as spirocyclic rings, bicyclic rings
and the like, which comprise a single heteroatom will be characterized and referred
to herein as being encompassed by the cyclic family corresponding to the heteroatom
containing ring, although the artisan can have alternative characterizations. For
example, 1,2,3,4-tetrahydroquinoline having the formula:
is, for the purposes of the present disclosure, considered a heterocyclic unit. 6,7-Dihydro-5
H-cyclopentapyrimidine having the formula:
is, for the purposes of the present disclosure, considered a heteroaryl unit. When
a fused ring unit contains heteroatoms in both a saturated ring (heterocyclic ring)
and an aryl ring (heteroaryl ring), the aryl ring will predominate and determine the
type of category to which the ring is assigned herein for the purposes of describing
the disclosure. For example, 1,2,3,4-tetrahydro-[1,8]naphthyridine having the formula:
is, for the purposes of the present disclosure, considered a heteroaryl unit.
[0044] The term "substituted" is used throughout the specification. The term "substituted"
is applied to the units described herein as "substituted unit or moiety is a hydrocarbyl
unit or moiety, whether acyclic or cyclic, which has one or more hydrogen atoms replaced
by a substituent or several substituents as defined herein below." The units, when
substituting for hydrogen atoms are capable of replacing one hydrogen atom, two hydrogen
atoms, or three hydrogen atoms of a hydrocarbyl moiety at a time. In addition, these
substituents can replace two hydrogen atoms on two adjacent carbons to form said substituent,
new moiety, or unit. For example, a substituted unit that requires a single hydrogen
atom replacement includes halogen, hydroxyl, and the like. A two hydrogen atom replacement
includes carbonyl, oximino, and the like. A two hydrogen atom replacement from adjacent
carbon atoms includes epoxy, and the like. Three hydrogen replacement includes cyano,
and the like. The term substituted is used throughout the present specification to
indicate that a hydrocarbyl moiety,
inter alia, aromatic ring, alkyl chain; can have one or more of the hydrogen atoms replaced by
a substituent. When a moiety is described as "substituted" any number of the hydrogen
atoms can be replaced. For example, 4-hydroxyphenyl is a "substituted aromatic carbocyclic
ring (aryl ring)", (N,N-dimethyl-5-amino)octanyl is a " substituted C
8 linear alkyl unit, 3-guanidinopropyl is a "substituted C
3 linear alkyl unit," and 2-carboxypyridinyl is a "substituted heteroaryl unit."
[0045] The following are non-limiting examples of units which can substitute for hydrogen
atoms on a carbocyclic, aryl, heterocyclic, or heteroaryl unit:
- i) substituted or unsubstituted C1-C12 linear, C3-C12 branched, or C3-C12 cyclic alkyl; for example, methyl (C1), chloromethyl (C1), trifluoromethyl (C1), aminomethyl (C1), ethyl (C2), hydroxymethyl 1-chloroethyl (C2), 2-hydroxyethyl (C2), 1,2-difluoroethyl (C2), n-propyl (C3), iso-propyl (C3), 3-carboxypropyl (C3), cyclopropyl (C3), 2-methyl-cyclopropyl (C3), n-butyl (C4), sec-butyl (C4), iso-butyl (C4), tert-butyl (C4), cyclobutyl (C4), 2,3-dihydroxycyclobutyl (C4), pentyl (C5), cyclopentyl (C5), hexyl (C6), and cyclohexyl (C6), and the like;
- ii) substituted or unsubstituted C2-C12 linear, C3-C12 branched, or C3-C12 cyclic alkenyl; for example, ethenyl (C2), 2-chloroethenyl (also 2-chlorovinyl) (C2), 3-propenyl (C3), 1-propenyl (also 2-methylethenyl) (C3), isopropenyl (also 2-methylethen-2-yl) (C3), buten-4-yl (C4), 4-hydroxybuten-1-yl (C4), cyclobutenyl (C4), cyclopentenyl (C5), cyclopentadienyl (C5), cyclohexenyl (C6), 7-hydroxy-7-methyloct-4-en-2-yl (C9), and 7-hydroxy-7-methyloct-3,5-dien-2-yl (C9), and the like;
- iii) substituted or unsubstituted C2-C12 linear or C3-C12 branched alkynyl; for example, ethynyl (C2), prop-2-ynyl (also propargyl) (C3), propyn-1-yl (C3), 2-methyl-hex-4-yn-1-yl (C7); 5-hydroxy-5-methylhex-3-ynyl (C7), 6-hydroxy-6-methylhept-3-yn-2-yl (C8), 5-hydroxy-5-ethylhept-3-ynyl (C9), and the like;
- iv) substituted or unsubstituted C6 or C10 aryl; for example, phenyl, 2-chlorophenyl, 3-hydroxyphenyl, 4-nitrophenyl, 2-fluoro-4-methylphenyl,
3,5-dinitrophenyl, 8-hydroxynaphth-1-yl, 6-sulfonylnapth-2-yl, and the like;
- v) substituted or unsubstituted C1-C9 heterocyclic; for example, as defined further herein;
- vi) substituted or unsubstituted C1-C11 heteroaryl; for example, as defined further herein;
- vii) halogen; for example, fluoro, chloro, bromo, and iodo;
- viii) -[C(R23a)(R23b)]xOR10;
R10 is chosen from:
- a) -H;
- b) substituted or unsubstituted C1-C12 linear, C3-C12 branched, or C3-C12 cyclic alkyl;
- c) C6 or C10 substituted or unsubstituted aryl or alkylenearyl;
- d) C1-C9 substituted or unsubstituted heterocyclic;
- e) C1-C11 substituted or unsubstituted heteroaryl;
- ix) -[C(R23a)(R23b)]xN(R11a)(R11b);
R11a and R11b are each independently chosen from:
- a) -H;
- b) -OR12;
R12 is hydrogen or C1-C4 linear alkyl;
- c) substituted or unsubstituted C1-C12 linear, C3-C12 branched, or C3-C12 cyclic alkyl;
- d) C6 or C10 substituted or unsubstituted aryl;
- e) C1-C9 substituted or unsubstituted heterocyclic;
- f) C1-C11 substituted or unsubstituted heteroaryl; or
- g) R11a and R11b can be taken together to form a substituted or unsubstituted ring having from 3 to
10 carbon atoms and from 0 to 3 heteroatoms chosen from oxygen, nitrogen, and sulfur;
- x) -[C(R23a)(R23b)]xC(O)R13;
R13 is:
- a) substituted or unsubstituted C1-C12 linear, C3-C12 branched, or C3-C12 cyclic alkyl;
- b) -OR14;
R14 is hydrogen, substituted or unsubstituted C1-C4 linear alkyl, C6 or C10 substituted or unsubstituted aryl, C1-C9 substituted or unsubstituted heterocyclic, C1-C11 substituted or unsubstituted heteroaryl;
- c) -N(R15a)(R15b);
R15a and R15b are each independently hydrogen, substituted or unsubstituted C1-C12 linear, C3-C12 branched, or C3-C12 cyclic alkyl; C6 or C10 substituted or unsubstituted aryl; C1-C9 substituted or unsubstituted heterocyclic; C1-C11 substituted or unsubstituted heteroaryl; or R15a and R15b can be taken together to form a substituted or unsubstituted ring having from 3 to
10 carbon atoms and from 0 to 3 heteroatoms chosen from oxygen, nitrogen, and sulfur;
- xi) -[C(R23a)(R23b)]xOC(O)R16;
R16 is:
- a) substituted or unsubstituted C1-C12 linear, C3-C12 branched, or C3-C12 cyclic alkyl;
- b) -N(R17a)(R17b);
R17a and R17b are each independently hydrogen, substituted or unsubstituted C1-C12 linear, C3-C12 branched, or C3-C12 cyclic alkyl; C6 or C10 substituted or unsubstituted aryl; C1-C9 substituted or unsubstituted heterocyclic; C1-C11 substituted or unsubstituted heteroaryl; or R17a and R17b can be taken together to form a substituted or unsubstituted ring having from 3 to
10 carbon atoms and from 0 to 3 heteroatoms chosen from oxygen, nitrogen, and sulfur;
- xii) -[C(R23a)(R23b)]xNR18C(O)R19;
R18 is:
- a) -H; or
- b) substituted or unsubstituted C1-C4 linear, C3-C4 branched, or C3-C4 cyclic alkyl;
R19 is:
- a) substituted or unsubstituted C1-C12 linear, C3-C12 branched, or C3-C12 cyclic alkyl;
- b) -N(R20a)(R20b);
R20a and R20b are each independently hydrogen, substituted or unsubstituted C1-C12 linear, C3-C12 branched, or C3-C12 cyclic alkyl; C6 or C10 substituted or unsubstituted aryl; C1-C9 substituted or unsubstituted heterocyclic; C1-C11 substituted or unsubstituted heteroaryl; or R20a and R20b can be taken together to form a substituted or unsubstituted ring having from 3 to
10 carbon atoms and from 0 to 3 heteroatoms chosen from oxygen, nitrogen, and sulfur;
- xiii) -[C(R23a)(R23b)]xCN;
- xiv) -[C(R23a)(R23b)]xNO2;
- xv) -[C(R23a)(R23b)]xR21;
R21 is C1-C10 linear, C3-C10 branched, or C3-C10 cyclic alkyl substituted by from 1 to 21 halogen atoms chosen from -F, -Cl, -Br,
or -I;
- xvi) -[C(R23a)(R23b)]xSO2R22;
R22 is hydrogen, hydroxyl, substituted or unsubstituted C1-C4 linear or C3-C4 branched alkyl; substituted or unsubstituted C6, C10, or C14 aryl; C7-C15 alkylenearyl; C1-C9 substituted or unsubstituted heterocyclic; or C1-C11 substituted or unsubstituted heteroaryl;
R
23a and R
23b are each independently hydrogen or C
1-C
4 alkyl; and
the index x is an integer from 0 to 5.
[0046] The compounds disclosed herein include all salt forms, for example, salts of both
basic groups,
inter alia, amines, as well as salts of acidic groups,
inter alia, carboxylic acids. The following are non-limiting examples of anions that can form
salts with basic groups: chloride, bromide, iodide, sulfate, bisulfate, carbonate,
bicarbonate, phosphate, formate, acetate, propionate, butyrate, pyruvate, lactate,
oxalate, malonate, maleate, succinate, tartrate, fumarate, citrate, and the like.
The following are non-limiting examples of cations that can form salts of acidic groups:
sodium, lithium, potassium, calcium, magnesium, bismuth, and the like.
[0047] For the purposes of the present disclosure the terms "compound," "analog," and "composition
of matter" stand equally well for one another and include all enantiomeric forms,
diastereomeric forms, salts, and the like, and the terms "compound," "analog," and
"composition of matter."
HIF-1α Prolyl Hydroxylase Inhibitors
[0048] The disclosed compounds have the following formulae:
wherein L is chosen from CH
2 or SO
2, thereby providing for
N-substituted benzyl or
N-substituted sulfonylaryl-3-hydroxypyridin-2-(1
H)-ones. Y, R
1 and R
2 are further defined herein below.
[0049] Disclosed herein are
N-substituted benzyl and
N-substituted sulfonylaryl-4-aminomethylene-3-hydroxypyridin-2-(1
H)-ones that are HIF-1α prolyl hydroxylase inhibitors having the formula:
wherein R
1 and R
2 are further defined herein below.
Alkyl piperizine-1-carboxylates
[0050] One category of these compounds relates to C
1-C
4 linear or branched alkyl 4-{[(1-
N-(chloro- or fluoro-substituted)-benzyl]-3-hydroxy-2-oxo-1,2-dihydropyridin-4-yl)methyl}piperazine-1-carboxylates
having the formula:
wherein Z is a phenyl group that is substituted with from 1 to 5 halogen atomsthat
are chosen from chloro and fluoro, and R
1 and R
2 are taken together to form a piperazine ring that is substituted with alkylcarboxy
unit wherein R
4 is chosen from C
1-C
4 linear or C
3-C
4 branched alkyl, for example,
tert butyl 4{[1-(4chlorobenzyl)-3-hydroxy-2-oxo 1,2-dihydropyridin-4-yl]methyl}piperazine-1-carboxylate
having the formula:
[0051] One aspect of R
4 units relates to compounds wherein R
4 is
tert-butyl (C
4). Another aspect of R
4 units relates to compounds wherein R
4 is methyl (C
1). A further aspect of R
4 units relates to compounds wherein R
4 is ethyl (C
2). A still further aspect of R
4 units relates to compounds wherein R
4 is chosen from n-propyl (C
3),
iso-propyl (C
3), n-butyl (C
4),
sec-butyl (C
4), and
iso-butyl (C
4). R
4 is not hydrogen, therefore, a carboxylate unit having the formula: -CO
2H is expressly excluded from this category, but may be included in other categories
as described herein below.
[0052] Z is phenyl substituted with from 1 to 5 halogens chosen from fluorine and chlorine.
One aspect of Z units relates to compounds wherein Z is 4-chlorophenyl. Another aspect
of Z units relates to compounds wherein Z is chosen from 2-chlorophenyl, 3-chlorophenyl,
2-fluorophenyl, 3-fluorophenyl, or 4-fluorophenyl. A further aspect of Z units relates
to compounds wherein Z is chosen from 2,3-difluorophenyl, 2,4-difluorophenyl, 2,5-difluorophenyl,
2,6-difluorophenyl, 2,3-dichlorophenyl, 2,4-dichlorophenyl, 2,5-dichlorophenyl, and
2,6-dichlorophenyl.
[0053] The following are non-limiting examples of compounds according to this category:
methyl 4-{[1-(4-chlorobenzyl)-3-hydroxy-2-oxo-1,2-dihydropyridin-4-yl]methyl}-piperazine-1-carboxylate
having the formula:
methyl 4-{[1-(3-chlorobenzyl)-3-hydroxy-2-oxo-1,2-dihydropyridin-4-yl]methyl}-piperazine-1-carboxylate
having the formula:
methyl 4-{[1-(2-chlorobenzyl)-3-hydroxy-2-oxo-1,2-dihydropyridin-4-yl]methyl}-piperazine-1-carboxylate
having the formula:
ethyl 4-{[1-(4-chlorobenzyl)-3-hydroxy-2-oxo-1,2-dihydropyridin-4-yl]methyl}-piperazine-1-carboxylate
having the formula:
ethyl 4-{[1-(3-chlorobenzyl)-3-hydroxy-2-oxo-1,2-dihydropyridin-4-yl]methyl}-piperazine-1-carboxylate
having the formula:
ethyl 4-{[1-(2-chlorobenzyl)-3-hydroxy-2-oxo-1,2-dihydropyridin-4-yl]methyl}-piperazine-1-carboxylate
having the formula:
tert-butyl 4-{[1-(4-chlorobenzyl)-3-hydroxy-2-oxo-1,2-dihydropyridin-4-yl]methyl}-piperazine-1-carboxylate
having the formula:
tert-butyl 4-{[1-(3-chlorobenzyl)-3-hydroxy-2-oxo-1,2-dihydropyridin-4-yl]methyl}-piperazine-1-carboxylate
having the formula:
tert-butyl 4-{[1-(2-chlorobenzyl)-3-hydroxy-2-oxo-1,2-dihydropyridin-4-yl]methyl}-piperazine-1-carboxylate
having the formula:
methyl 4-{[1-(4-fluorobenzyl)-3-hydroxy-2-oxo-1,2-dihydropyridin-4-yl]methyl}-piperazine-1-carboxylate
having the formula:
methyl 4-{[1-(3-fluorobenzyl)-3-hydroxy-2-oxo-1,2-dihydropyridin-4-yl]methyl}-piperazine-1-carboxylate
having the formula:
methyl 4-{[1-(2-fluorobenzyl)-3-hydroxy-2-oxo-1,2-dihydropyridin-4-yl]methyl}-piperazine-1-carboxylate
having the formula:
ethyl 4- {[1-(4-fluorobenzyl)-3-hydroxy-2-oxo-1,2-dihydropyridin-4-yl]methyl}-piperazine-1-carboxylate
having the formula:
ethyl 4-{[1-(3-fluorobenzyl)-3-hydroxy-2-oxo-1,2-dihydropyridin-4-yl]methyl}-piperazine-1-carboxylate
having the formula:
ethyl 4-{[1-(2-fluorobenzyl)-3-hydroxy-2-oxo-1,2-dihydropyridin-4-yl]methyl}-piperazine-1-carboxylate
having the formula:
tert-butyl 4-{[1-(4-fluorobenzyl)-3-hydroxy-2-oxo-1,2-dihydropyridin-4-yl]methyl}-piperazine-1-carboxylate
having the formula:
tert-Butyl 4- {[1-(3-fluorobenzyl)-3-hydroxy-2-oxo-1,2-dihydropyridin-4-yl]methyl}-piperazine-1-carboxylate
having the formula:
and
tert-butyl 4- {[1-(2-fluorobenzyl)-3-hydroxy-2-oxo-1,2-dihydropyridin-4-yl]methyl}piperazine-1-carboxylate
having the formula:
[0054] Another category of compounds relates to
N-unsubstituted-benzyl-4-aminomethyl-3-hydroxypyridin-2-(1
H)-ones, wherein Z is an unsubstituted phenyl group, having the formula:
wherein R
1 and R
2 are are be taken together to form a substituted or unsubstituted heterocyclic or
heteroaryl ring.
[0055] A first aspect of this category relates to compounds having the formula:
wherein R
1 and R
2 are be taken together to form a substituted or unsubstituted heterocyclic or heteroaryl
ring represented by ring A having from 2 to 20 carbon atoms and from 1 to 7 heteroatoms,
and R
200 represents from 0 to 40 substitutions form hydrogen. The index w is an integer from
0 to 40. Non-limiting examples of rings include diazirinyl (C
1), 1,2,3,4-tetrazolyl (C
1), aziridinyl (C
2), urazolyl (C
2), [1,2,3]triazolyl (C
2), [1,2,4]triazolyl (C
2), azetidinyl (C
3), pyrazolidinyl (C
3), imidazolidinyl (C
3), oxazolidinyl (C
3), isoxazolinyl (C
3), isoxazolyl (C
3), thiazolidinyl (C
3), isothiazolyl (C
3), isothiazolinyl (C
3), oxathiazolidinonyl (C
3), oxazolidinonyl (C
3), hydantoinyl (C
3), 1
H-imidazolyl (C
3), pyrrolidinyl (C
4), morpholinyl (C
4), piperazinyl (C
4), piperidinyl (C
4), piperidin-2-onyl (valerolactam) (C
5), 7
H-purinyl (C
5), 9
H-purinyl (C
5), 6-amino-9
H-purinyl (C
5), 2,3,4,5-tetrahydro-1
H-azepinyl (C
6), 5
H-pyrrolo[3,2-
d]pyrimidinyl (C
6), 7
H-pyrrolo[2,3-d]pyrimidinyl (C
6), and 1,2,3,4-tetrahydroquinoline (C
9).
[0056] Each R
200 unit is independently chosen from:
i) substituted or unsubstituted C1-C12 linear, C3-C12 branched, or C3-C12 cyclic alkyl; for example, methyl (C1), (C1), chloromethyl (C1), trifluoromethyl (C1), aminomethyl (C1), ethyl (C2), hydroxymethyl 1-chloroethyl (C2), 2-hydroxyethyl (C2), 1,2-difluoroethyl (C2), n-propyl (C3), iso-propyl (C3), 3-carboxypropyl (C3), cyclopropyl (C3), 2-methyl-cyclopropyl (C3), n-butyl (C4), sec-butyl (C4), iso-butyl (C4), tert-butyl (C4), cyclobutyl (C4), 2,3-dihydroxycyclobutyl (C4), pentyl (C5), cyclopentyl (C5), hexyl (C6), and cyclohexyl (C6), and the like;
ii) substituted or unsubstituted C2-C12 linear, C3-C12 branched, or C3-C12 cyclic alkenyl; for example, ethenyl (C2), 2-chloroethenyl (also 2-chlorovinyl) (C2), 3-propenyl (C3), 1-propenyl (also 2-methylethenyl) (C3), isopropenyl (also 2-methylethen-2-yl) (C3), buten-4-yl (C4), 4-hydroxybuten-1-yl (C4), cyclobutenyl (C4), cyclopentenyl (C5), cyclopentadienyl (C5), cyclohexenyl (C6), 7-hydroxy-7-methyloct-4-en-2-yl (C9), and 7-hydroxy-7-methyloct-3,5-dien-2-yl (C9), and the like;
iii) substituted or unsubstituted C1-C12 linear or C3-C12 branched alkynyl; for example, ethynyl (C2), prop-2-ynyl (also propargyl) (C3), propyn-1-yl (C3), 2-methyl-hex-4-yn-1-yl (C7); 5-hydroxy-5-methylhex-3-ynyl (C7), 6-hydroxy-6-methylhept-3-yn-2-yl (C8), 5-hydroxy-5-ethylhept-3-ynyl (C9), and the like;
iv) substituted or unsubstituted C6 or C10 aryl; for example, phenyl (C6), naphthylen-1-yl (C10), naphthylen-2-yl (C10), 4-fluorophenyl (C6), 2-hydroxyphenyl (C6), 3-methylphenyl (C6), 2-amino-4-fluorophenyl (C6), 2-(N,N-diethylamino)phenyl (C6), 2-cyanophenyl (C6), 2,6-di-tert-butylphenyl (C6), 3-methoxyphenyl (C6), 8-hydroxynaphthylen-2-yl (C10), 4,5-dimethoxynaphthylen-1-yl (C10), 6-cyano-naphthylen-1-yl (C10), and the like;
v) substituted or unsubstituted C1-C9 heterocyclic; for example, diazirinyl (C1), aziridinyl (C2), urazolyl (C2), azetidinyl (C3), pyrazolidinyl (C3), imidazolidinyl (C3), oxazolidinyl (C3), isoxazolinyl (C3), isoxazolyl (C3), thiazolidinyl (C3), isothiazolyl (C3), isothiazolinyl (C3), oxathiazolidinonyl (C3), oxazolidinonyl (C3), hydantoinyl (C3), tetrahydrofuranyl (C4), pyrrolidinyl (C4), morpholinyl (C4), piperazinyl (C4),piperidinyl (C4),dihydropyranyl (C5), tetrahydropyranyl (C5), piperidin-2-onyl (valerolactam) (C5), and the like;
vi) substituted or unsubstituted C1-C11 heteroaryl; for example, 1,2,3,4-tetrazolyl (C1), [1,2,3]triazolyl (C4),[1,2,4]triazolyl (C2), triazinyl (C3), thiazolyl (C3), 1H-imidazolyl (C3), oxazolyl (C3), furanyl (C4),thiopheneyl (C4),pyrimidinyl (C4), pyridinyl (C5), and the like;
vii) halogen; for example, -F, -Cl, -Br, or -I;
viii) -[C(R37a)(R37b)]yOR24;
R24 is chosen from:
- a) -H;
- b) substituted or unsubstituted C2-C12 linear, C3-C12 branched, or C3-C12 cyclic alkyl;
- c) substituted or unsubstituted C6 or C10 aryl or C7 or C10 alkylenearyl; for example, phenyl or benzyl
- d) substituted or unsubstituted C1-C9 heterocyclic;
- e) substituted or unsubstituted C1-C11 heteroaryl;
for example, -OH, -CH2OH, -OCH3, -CH2OCH3, -OCH2CH3, -CH2OCH2CH3, -OCH2CH2CH3, and -CH2OCH2CH2CH3;
ix) -[C(R37a)(R37b)]yN(R25a)(R25b);
R25a and R25b are each independently chosen from:
- a) -H;
- b) -OR26;
R26 is hydrogen or C1-C4 linear alkyl;
- c) substituted or unsubstituted C2-C12 linear, C3-C12 branched, or C3-C12 cyclic alkyl;
- d) substituted or unsubstituted C6 or C10 aryl;
- e) substituted or unsubstituted C1-C9 heterocyclic;
- f) substituted or unsubstituted C1-C11 heteroaryl; or
- g) R25a and R25b can be taken together to form a substituted or unsubstituted ring having from 3 to
10 carbon atoms and from 0 to 3 heteroatoms chosen from oxygen, nitrogen, and sulfur;
for example, -NH2, -CH2NH2, -NHCH3, -N(CH3)2, NHOH, -NHOCH3, -NH(CH2CH3), -CH2NHCH3, -CH2N(CH3)2, -CH2NH(CH2CH3), and the like;
x) -[C(R37a)(R37b)]yC(O)R27;
R21 is:
- a) substituted or unsubstituted C2-C12 linear, C3-C12 branched, or C3-C12 cyclic alkyl;
- b) -OR28;
R28 is hydrogen, substituted or unsubstituted C1-C4 linear alkyl, substituted or unsubstituted C6 or C10 aryl, substituted or unsubstituted C1-C9 heterocyclic, substituted or unsubstituted C1-C11 heteroaryl;
- c) N(R29a)(R29b);
R29a and R29b are each independently hydrogen, substituted or unsubstituted C2-C12 linear, C3-C12 branched, or C3-C12 cyclic alkyl; substituted or unsubstituted C6 or C10 aryl, substituted or unsubstituted C1-C9 heterocyclic, substituted or unsubstituted C1-C11 heteroaryl; or R29a and R29b can be taken together to form a substituted or unsubstituted ring having from 3 to
10 carbon atoms and from 0 to 3 heteroatoms chosen from oxygen, nitrogen, and sulfur;
for example, -COCH3, -CH2COCH3, -OCH2CH3, -CH2COCH2CH3, -COCH2CH2CH3, -CH2COCH2CH2CH3, and the like;
xi) -[C(R37a)(R37b)]yOC(O)R30;
R30 is:
- a) C1-C12 substituted or unsubstituted linear, branched, or cyclic alkyl;
- b) -N(R31a)(R31b);
R31a and R31b are each independently hydrogen, substituted or unsubstituted C2-C12 linear, C3-C12 branched, or C3-C12 cyclic alkyl; substituted or unsubstituted C6 or C10 aryl, substituted or unsubstituted C1-C9 heterocyclic, substituted or unsubstituted C1-C11 heteroaryl; or R31a and R31b can be taken together to form a substituted or unsubstituted ring having from 3 to
10 carbon atoms and from 0 to 3 heteroatoms chosen from oxygen, nitrogen, and sulfur;
for example, -OC(O)CH3, -CH2OC(O)CH3, -OC(O)NH2. -CH2OC(O)NH2, -OC(O)NHCH3, -CH2OC(O)NHCH3, -OC(O)N(CH3)2, -CH2OC(O)N(CH3)2, and the like;
xii) -[C(R37a)(R37b)]yNR32C(O)R33;
R32 is:
- a) -H; or
- b) substituted or unsubstituted C1-C4 linear, C3-C4 branched, or C3-C4 cyclic alkyl;
R
33is:
- a) substituted or unsubstituted C2-C12 linear, C3-C12 branched, or C3-C12 cyclic alkyl;
- b) -N(R34a)(R34b);
R34a and R34b are each independently hydrogen, substituted or unsubstituted C2-C12 linear, C3-C12 branched, or C3-C12 cyclic alkyl; substituted or unsubstituted C6 or C10 aryl, substituted or unsubstituted C1-C9 heterocyclic, substituted or unsubstituted C1-C11 heteroaryl; C1-C11 substituted or unsubstituted heteroaryl; or R34a and R34b can be taken together to form a substituted or unsubstituted ring having from 3 to
10 carbon atoms and from 0 to 3 heteroatoms chosen from oxygen, nitrogen, and sulfur;
for example, -NHC(O)CH3, -CH2NHC(O)CH3, -NHC(O)NH2, -CH2NHC(O)NH2, -NHC(O)NHCH3, -CH2NHC(O)NHCH3, -OC(O)N(CH3)2, -CH2NHC(O)N(CH3)2, and the like;
xiii) -[C(R37a)(R37b)]yCN; for example; -CN, -CH2CN, and -CH2CH2CN;
xiv) -[C(R37a)(R37b)]yNO2; for example; -NO2, -CH2NO2, and -CH2CH2NO2;
xv) -[C(R37a)(R37b)]yR35; for example, -CH2F, -CHF2, -CF3, -CCl3, or -CBr3; R35 is C1-C10 linear, C3-C10 branched, or C3-C10 cyclic alkyl substituted by from 1 to 21 halogen atoms chosen from -F, -Cl, -Br,
or -I;
xvi) -[C(R37a)(R37b)]ySO2R36;
R36 is hydrogen, hydroxyl, substituted or unsubstituted C1-C4 linear or C3-C4 branched alkyl; substituted or unsubstituted C6, C10, or C14 aryl; C7-C15 alkylenearyl; substituted or unsubstituted C1-C9 heterocyclic; or substituted or unsubstituted C1-C11 heteroaryl;
for example, -SO2H, -CH2SO2H, -SO2CH3, -CH2SO2CH3, -SO2C6H5, and -CH2SO2C6H5; and
xv) two hydrogen atoms on a ring carbon atom can be substituted to form a =O, =S,
or =NH unit;
R
37a and R
37b are each independently hydrogen or C
1-C
4 alkyl; and
the index y is an integer from 0 to 5.
[0057] A first embodiment of this aspect relates to compounds wherein R
1 and R
2 are taken together to form a 5-member substituted or unsubstituted C
1-C
4 heterocyclic or a substituted or unsubstituted C
1-C
4 heteroaryl ring, non-limiting examples of which include a ring chosen from:
i)
ii)
ii)
iii)
iv)
v)
vi)
vii)
or
viii)
[0058] A first iteration of this embodiment relates to HIF-1α prolyl hydroxylase inhibitors
having the formula:
R
200 represents from 0 to 2 substitutions for a ring hydrogen, wherein the substitutions
for hydrogen are independently chosen from:
i) C1-C4 linear or C3-C4 branched alkyl;
ii) C1-C4 linear or C3-C4 branched alkoxy;
iii) hydroxyl;
iv) cyano;
v) nitro;
vi) amino, methylamino, or dimethylamino;
vii) carboxy, methyl carboxy; or ethyl carboxy;
viii) formyl, acetyl, or propionyl;
ix) amido, methyl amido, or dimethyl amido;
x) halogen;
xi) heterocyclic; or
xii) heteroaryl.
[0059] Non-limiting examples of this iteration include HIF-1α prolyl hydroxylase inhibitors
having the formula:
[0060] A further iteration of this embodiment relates to HIF-1α prolyl hydroxylase inhibitors
wherein R
1 and R
2 are taken together to form a 5-member substituted or unsubstituted heterocyclic or
heteroaryl ring having more than one heteroatom in the ring. Non-limiting examples
include:
[0061] Another embodiment of this aspect relates to HIF-1α prolyl hydroxylases inhibitors
wherein R
1 and R
2 are taken together to form a substituted or unsubstituted C
4-C
11 heterocyclic or a substituted or unsubstituted C
4-C
11 heteroaryl ring, non-limiting examples of which are chosen from:
- i)
- ii)
- iii)
- iv)
or
- v)
[0063] Another category of compounds has the formula:
wherein R
200 and the index w are the same as defined herein above. R represents from 0 to 5 substitutions
for hydrogen, wherein each R is independently chosen from:
- i) C1-C12 substituted or unsubstituted linear, branched, or cyclic alkyl; for example, methyl
(C1), (C1), chloromethyl (C1), trifluoromethyl (C1), aminomethyl (C1), ethyl (C4),hydroxymethyl 1-chloroethyl (C2), 2-hydroxyethyl (C2), 1,2-difluoroethyl (C2), n-propyl (C3), iso-propyl (C3), 3-carboxypropyl (C3), cyclopropyl (C3), 2-methyl-cyclopropyl (C3), n-butyl (C4),sec-butyl (C4), iso-butyl(C4), tert-butyl (C4), cyclobutyl (C4),2,3-dihydroxycyclobutyl (C4), pentyl (C5), cyclopentyl (C5), hexyl (C6), and cyclohexyl (C6), and the like;
- ii) C2-C12 substituted or unsubstituted linear, branched, or cyclic alkenyl; for example, ethenyl
(C4),2-chloroethenyl (also 2-chlorovinyl) (C2), 3-propenyl (C3), 1-propenyl (also 2-methylethenyl) (C3), isopropenyl (also 2-methylethen-2-yl) (C3), buten-4-yl (C4),4-hydroxybuten-1-yl (C4), cyclobutenyl (C4), cyclopentenyl (C5), cyclopentadienyl (C5), cyclohexenyl (C6), 7-hydroxy-7-methyloct-4-en-2-yl (C9), and 7-hydroxy-7-methyloct-3,5-dien-2-yl (C9), and the like;
- iii) C2-C12 substituted or unsubstituted linear or branched alkynyl; for example, ethynyl (C2), prop-2-ynyl (also propargyl) (C3), propyn-1-yl (C3), 2-methyl-hex-4-yn-1-yl (C7); 5-hydroxy-5-methylhex-3-ynyl (C7), 6-hydroxy-6-methylhept-3-yn-2-yl (C8), 5-hydroxy-5-ethylhept-3-ynyl (C9), and the like;
- iv) C6 or C10 substituted or unsubstituted aryl; for example, phenyl (C6), naphthylen-1-yl (C10), naphthylen-2-yl (C10), 4-fluorophenyl (C6), 2-hydroxyphenyl (C6), 3-methylphenyl (C6), 2-amino-4-fluorophenyl (C6), 2-(N,N-diethylamino)phenyl (C6), 2-cyanophenyl (C6), 2,6-di-tert-butylphenyl (C6), 3-methoxyphenyl (C6), 8-hydroxynaphthylen-2-yl (C10), 4,5-dimethoxynaphthylen-1-yl (C10), 6-cyano-naphthylen-1-yl (C10), and the like;
- v) C1-C9 substituted or unsubstituted heterocyclic; for example, diazirinyl (C1), aziridinyl (C4),urazolyl (C2), azetidinyl (C3), pyrazolidinyl (C3), imidazolidinyl (C3), oxazolidinyl (C3), isoxazolinyl (C3), isoxazolyl (C3), thiazolidinyl (C3), isothiazolyl (C3), isothiazolinyl (C3), oxathiazolidinonyl (C3), oxazolidinonyl (C3), hydantoinyl (C3), tetrahydrofuranyl (C4), pyrrolidinyl (C4), morpholinyl (C4), piperazinyl (C4), piperidinyl (C4), dihydropyranyl (C5), tetrahydropyranyl (C5), piperidin-2-onyl (valerolactam) (C5), and the like;
- vi) C1-C11 substituted or unsubstituted heteroaryl; for example, 1,2,3,4-tetrazolyl (C1), [1,2,3]triazolyl (C2), [1,2,4]triazolyl (C2), triazinyl (C3), thiazolyl (C3), 1H-imidazolyl (C3), oxazolyl (C3), furanyl (C4), thiopheneyl (C4), pyrimidinyl (C4), pyridinyl (C5), and the like;
- vii) halogen; for example, -F, -Cl, -Br, or -I;
- viii) -[C(R23a)(R23b)]xOR10;
R10 is chosen from:
- a) -H;
- b) C1-C12 substituted or unsubstituted linear, branched, or cyclic alkyl;
- c) C6 or C10 substituted or unsubstituted aryl or alkylenearyl;
- d) C1-C9 substituted or unsubstituted heterocyclic;
- e) C1-C11 substituted or unsubstituted heteroaryl;
for example, -OH, -CH2OH, -OCH3, -CH2OCH3, -OCH2CH3, -CH2OCH2CH3, -OCH2CH2CH3, and -CH2OCH2CH2CH3;
- ix) -[C(R23a)(R23b)]xN(R11a)(R11b);
R11a and R11b are each independently chosen from:
- a) -H;
- b) -OR12;
R12 is hydrogen or C1-C4 linear alkyl;
- c) C1-C12 substituted or unsubstituted linear, branched, or cyclic alkyl;
- d) C6 or C10 substituted or unsubstituted aryl;
- e) C1-C9 substituted or unsubstituted heterocyclic;
- f) C1-C11 substituted or unsubstituted heteroaryl; or
- g) R11a and R11b can be taken together to form a substituted or unsubstituted ring having from 3 to
10 carbon atoms and from 0 to 3 heteroatoms chosen from oxygen, nitrogen, and sulfur;
for example, -NH2, -CH2NH2, -NHCH3, -N(CH3)2, -NHOH, -NHOCH3, -NH(CH2CH3), -CH2NHCH3, -CH2N(CH3)2, -CH2NH(CH2CH3), and the like;
- x) -[C(R23a)(R23b)]xC(O)R13;
R13 is:
- a) C1-C12 substituted or unsubstituted linear, branched, or cyclic alkyl;
- b) -OR14;
R14 is hydrogen, substituted or unsubstituted C1-C4 linear alkyl, C6 or C10 substituted or unsubstituted aryl, C1-C9 substituted or unsubstituted heterocyclic, C1-C11 substituted or unsubstituted heteroaryl;
- c) -N(R15a)(R15b);
R15a and R15b are each independently hydrogen, C1-C12 substituted or unsubstituted linear, branched, or cyclic alkyl; C6 or C10 substituted or unsubstituted aryl; C1-C9 substituted or unsubstituted heterocyclic; C1-C11 substituted or unsubstituted heteroaryl; or R15a and R15b, can be taken together to form a substituted or unsubstituted ring having from 3
to 10 carbon atoms and from 0 to 3 heteroatoms chosen from oxygen, nitrogen, and sulfur;
for example, -COCH3, -CH2COCH3, -OCH2CH3, -CH2COCH2CH3, -COCH2CH2CH3, -CH2COCH2CH2CH3, and the like;
- xi) -[C(R23a)(R23b)]XOC(O)R16;
R16 is:
- a) C1-C12 substituted or unsubstituted linear, branched, or cyclic alkyl;
- b) -N(R17a)(R17b);
R17a and R17b are each independently hydrogen, C1-C12 substituted or unsubstituted linear, branched, or cyclic alkyl; C6 or C10 substituted or unsubstituted aryl; C1-C9 substituted or unsubstituted heterocyclic; C1-C11 substituted or unsubstituted heteroaryl; or R17a and R17b can be taken together to form a substituted or unsubstituted ring having from 3 to
10 carbon atoms
and from 0 to 3 heteroatoms chosen from oxygen, nitrogen, and sulfur;
- xii) -[C(R23a)(R23b)]xNR18C(O)R19;
R18 is:
- a) -H; or
- b) C1-C4 substituted or unsubstituted linear, branched, or cyclic alkyl;
R19 is:
- a) C1-C12 substituted or unsubstituted linear, branched, or cyclic alkyl;
- b) -N(R20a)(R20b);
R20a and R20b are each independently hydrogen, C1-C12 substituted or unsubstituted linear, branched, or cyclic alkyl; C6 or C10 substituted or unsubstituted aryl; C1-C9 substituted or unsubstituted heterocyclic; C1-C11 substituted or unsubstituted heteroaryl; or R20a and R20b can be taken together to form a substituted or unsubstituted ring having from 3 to
10 carbon atoms and from 0 to 3 heteroatoms chosen from oxygen, nitrogen, and sulfur;
for example, -NHC(O)CH3, -CH2NHC(O)CH3, -NHC(O)NH2, -CH2NHC(O)NH2, -NHC(O)NHCH3, -CH2NHC(O)NHCH3, - OC(O)N(CH3)2, -CH2NHC(O)N(CH3)2, and the like;
- xiii) -[C(R23a)(R23b)]xCN; for example; -CN, -CH2CN, and -CH2CH2CN;
- xiv) -[C(R23a)(R23b)]xNO2; for example; -NO2, -CH2NO2, and -CH2CH2NO2;
- xv) -[C(R23a)(R23b)]xR21; for example, -CH2F, -CHF2, -CF3, -CCl3, or -CBr3;
R21 is C1-C10 linear, branched, or cyclic alkyl substituted by from 1 to 21 halogen atoms chosen
from -F, -Cl, -Br, or -I;
- xvi) -[C(R23a)(R23b)]xSO2R22;
R22 is hydrogen, hydroxyl, substituted or unsubstituted C1-C4 linear or branched alkyl; substituted or unsubstituted C6, C10, or C14 aryl; C7-C15 alkylenearyl; C1-C9 substituted or unsubstituted heterocyclic; or C1-C11 substituted or unsubstituted heteroaryl; for example, -SO2H, -CH2SO2H, -SO2CH3, -CH2SO2CH3, -SO2C6H5, and -CH2SO2C6H5;
R
23a and R
23b are each independently hydrogen or C
1-C
4 alkyl; and
the index x is an integer from 0 to 5.
[0064] Non-limiting examples of this category include compounds having the formula:
[0065] A further category of compounds relates to unsubstituted
N-benzyl-4-aminomethyl-3-hydroxypyridin-2-(1
H)-ones having the formula:
wherein R
1 and R
2 are each independently chosen from:
- i) hydrogen;
- ii) substituted or unsubstituted C1-C10 linear, branched, or cyclic alkyl;
- iii) substituted or unsubstituted C2-C10 linear, branched, or cyclic alkenyl;
- iv) substituted or unsubstituted C2-C10 linear or branched alkynyl;
- v) substituted or unsubstituted C6 or C10 aryl;
- vi) substituted or unsubstituted C1-C9 heterocyclic; or
- vii) substituted or unsubstituted C1-C9 heteroaryl.
[0066] The first aspect of this category relates to HIF-1α prolyl hydroxylase inhibitors
wherein R
2 is hydrogen and R
1 is substituted or unsubstituted C
1-C
9 heterocyclic or C
1-C
9 heteroaryl. In a first embodiment, R
1 is a substituted heterocyclic group, non-limiting examples of which include aziridinyl
(C
2), azetidinyl (C
3), pyrrolidinyl (C
4), morpholinyl (C
4), piperazinyl (C
4), piperidinyl (C
4), piperidin-2-onyl (valerolactam) (C
5), and azepan-2-only (caprolactam) (C
6), wherein the R
1 unit can be bonded to the nitrogen atom at any position in the ring. In addition,
the C
1-C
9 heterocyclic or C
1-C
9 heteroaryl ring can be substituted at any position whether a ring carbon or a ring
heteroatom, for example, a ring nitrogen. Non-limiting examples of this embodiment
include:
[0067] In another embodiment, R
2 is hydrogen and R
1 is substituted or unsubstituted C
3-C
12 cycloalkyl wherein the cycloalkyl ring can be substituted at any ring position. Non-limiting
examples of this embodiment include:
[0068] A yet further category of compounds relates to unsubstituted
N-benzyl-4-aminomethyl-3-hydroxypyridin-2-(1
H)-ones having the formula:
R
1 and R
2 are each independently hydrogen or substituted or unsubstituted C
1-C
10 linear or branched alkyl, wherein the alkyl unit can be substituted by one or more
units independently chosen from:
- i) C1-C8 linear, branched, or cyclic alkoxy;
- ii) hydroxy;
- iii) halogen;
- iv) cyano;
- v) amino, C1-C8 mono-alkylamino, C1-C8 di-alkylamino;
- vi) -SR40; R40 is hydrogen or C1-C4 linear or branched alkyl;
- vii) substituted or unsubstituted C6 of C10 aryl;
- viii) substituted or unsubstituted C1-C9 heterocyclic; or
- ix) substituted or unsubstituted C1-C9 heteroaryl.
[0069] Non-limiting examples of this category include:
[0070] A still further category of the disclosed compounds has the formula:
wherein R
200 and the index w are the same as defined herein above. R represents from 0 to 5 substitutions
for hydrogen, wherein each R is independently chosen from:
i) substituted or unsubstituted C1-C12 linear, C3-C12 branched, or C3-C12 cyclic alkyl; for example, methyl (C1), (C1), chloromethyl (C1), trifluoromethyl (C2), aminomethyl (C1), ethyl (C2), hydroxymethyl 1-chloroethyl (C2), 2-hydroxyethyl (C2), 1,2-difluoroethyl (C2), n-propyl (C3), iso-propyl (C3), 3-carboxypropyl (C3), cyclopropyl (C3), 2-methyl-cyclopropyl (C3), n-butyl (C4), sec-butyl (C4), iso-butyl (C4), tert-butyl (C4), cyclobutyl (C4), 2,3-dihydroxycyclobutyl (C4), pentyl (C5), cyclopentyl (C5), hexyl (C6), and cyclohexyl (C6), and the like;
ii) substituted or unsubstituted C2-C12 linear, C3-C12 branched, or C3-C12 cyclic alkenyl; for example, ethenyl (C2), 2-chloroethenyl (also 2-chlorovinyl) (C2), 3-propenyl (C3), 1-propenyl (also 2-methylethenyl) (C3), isopropenyl (also 2-methylethen-2-yl) (C3), buten-4-yl (C4), 4-hydroxybuten-1-yl (C4), cyclobutenyl (C4), cyclopentenyl (C5), cyclopentadienyl (C5), cyclohexenyl (C6), 7-hydroxy-7-methyloct-4-en-2-yl (C9), and 7-hydroxy-7-methyloct-3,5-dien-2-yl (C9), and the like;
iii) substituted or unsubstituted C2-C12 linear or C3-C12 branched alkynyl; for example, ethynyl (C2), prop-2-ynyl (also propargyl) (C3), propyn-1-yl (C3), 2-methyl-hex-4-yn-1-yl (C7); 5-hydroxy-5-methylhex-3-ynyl (C7), 6-hydroxy-6-methylhept-3-yn-2-yl (C8), 5-hydroxy-5-ethylhept-3-ynyl (C9), and the like;
iv) substituted or unsubstituted C6 or C10 aryl; for example, phenyl (C6), naphthylen-1-yl (C10), naphthylen-2-yl (C10), 4-fluorophenyl (C6), 2-hydroxyphenyl (C6), 3-methylphenyl (C6), 2-amino-4-fluorophenyl (C6), 2-(N,N-diethylamino)phenyl (C6), 2-cyanophenyl (C6), 2,6-di-tert-butylphenyl (C6), 3-methoxyphenyl (C6), 8-hydroxynaphthylen-2-yl (C10), 4,5-dimethoxynaphthylen-1-yl (C10), 6-cyano-naphthylen-1-yl (C10), and the like;
v) substituted or unsubstituted C1-C9 heterocyclic; for example, diazirinyl (C1), aziridinyl (C2), urazolyl (C2), azetidinyl (C3), pyrazolidinyl (C3), imidazolidinyl (C3), oxazolidinyl (C3), isoxazolinyl (C3), isoxazolyl (C3), thiazolidinyl (C3), isothiazolyl (C3), isothiazolinyl (C3), oxathiazolidinonyl (C3), oxazolidinonyl (C3), hydantoinyl (C3), tetrahydrofuranyl (C4), pyrrolidinyl (C4), morpholinyl (C4), piperazinyl (C4), piperidinyl (C4), dihydropyranyl (C5), tetrahydropyranyl (C5), piperidin-2-onyl (valerolactam) (C5), and the like;
vi) substituted or unsubstituted C1-C11 heteroaryl; for example, 1,2,3,4-tetrazolyl (C1), [1,2,3]triazolyl (C2), [1,2,4]triazolyl (C2), triazinyl (C3), thiazolyl (C3), 1H-imidazolyl (C3), oxazolyl (C3), furanyl (C4), thiopheneyl (C4), pyrimidinyl (C4), pyridinyl (C5), and the like;
vii) halogen; for example, -F, -Cl, -Br, or -I;
viii) -[C(R23a)(R23b)]xOR10;
R10 is chosen from:
- a) -H;
- b) substituted or unsubstituted C1-C12 linear, C3-C12 branched, or C3-C12 cyclic alkyl;
- c) substituted or unsubstituted C6 or C10 aryl or C7 or C10 alkylenearyl;
- d) substituted or unsubstituted C1-C9 heterocyclic;
- e) substituted or unsubstituted C1-C11 heteroaryl;
for example, -OH, -CH2OH, -OCH3, -CH2OCH3, -OCH2CH3, -CH2OCH2CH3, -OCH2CH2CH3, and -CH2OCH2CH2CH3;
ix) -[C(R23a)(R23b)]xN(R11a)(R11b);
R11a and R11b are each independently chosen from:
- a) -H;
- b) -OR12;
R12 is hydrogen or C1-C4 linear alkyl;
- c) substituted or unsubstituted C1-C12 linear, C3-C12 branched, or C3-C12 cyclic alkyl;
- d) substituted or unsubstituted C6 or C10 aryl;
- e) substituted or unsubstituted C1-C9 heterocyclic;
- f) substituted or unsubstituted C1-C11 heteroaryl; or
- g) R11a and R11b can be taken together to form a substituted or unsubstituted ring having from 3 to
10 carbon atoms and from 0 to 3 heteroatoms chosen from oxygen, nitrogen, and sulfur;
for example, -NH2, -CH2NH2, -NHCH3, -N(CH3)2, NHOH, -NHOCH3, -NH(CH2CH3), -CH2NHCH3, -CH2N(CH3)2, -CH2NH(CH2CH3), and the like;
x) -[C(R23a)(R23b)]xC(O)R13;
R13 is:
- a) substituted or unsubstituted C1-C12 linear, C3-C12 branched, or C1-C12 cyclic alkyl;
- b) -OR14;
R14 is hydrogen, substituted or unsubstituted C1-C4 linear alkyl, substituted or unsubstituted C6 or C10 aryl, substituted or unsubstituted C1-C9 heterocyclic, substituted or unsubstituted C1-C11 heteroaryl;
- c) -N(R15a)(R15b);
R15a and R15b are each independently hydrogen, substituted or unsubstituted C1-C12 linear, C3-C12 branched, or C3-C12 cyclic alkyl; substituted or unsubstituted C6 or C10 aryl; substituted or unsubstituted C1-C9 heterocyclic; substituted or unsubstituted C1-C11 heteroaryl; or R15a and R15b can be taken together to form a substituted or unsubstituted ring having from 3 to
10 carbon atoms and from 0 to 3 heteroatoms chosen from oxygen, nitrogen, and sulfur;
for example, -COCH3, -CH2COCH3, -OCH2CH3, -CH2COCH2CH3, -COCH2CH2CH3, -CH2COCH2CH2CH3, and the like;
xi) -[C(R23a)(R23b)]xOC(O)R16;
R16 is:
- a) substituted or unsubstituted C1-C12 linear, C3-C12 branched, or C3-C12 cyclic alkyl;
- b) -N(R17a)(R17b);
R17a and R17b are each independently hydrogen, substituted or unsubstituted C1-C12 linear, C3-C12 branched, or C3-C12 cyclic alkyl; substituted or unsubstituted C6 or C10 aryl; substituted or unsubstituted C1-C9 heterocyclic; substituted or unsubstituted C1-C11 heteroaryl; or R17a and R17b can be taken together to form a substituted or unsubstituted ring having from 3 to
10 carbon atoms and from 0 to 3 heteroatoms chosen from oxygen, nitrogen, and sulfur;
xii) -[C(R23a)(R23b)]xNR18C(O)R19;
R18 is:
- a) -H; or
- b) substituted or unsubstituted C1-C4 linear, C3-C4 branched, or C3-C4 cyclic alkyl;
R
19is:
- a) substituted or unsubstituted C1-C12 linear, C3-C12 branched, or C3-C12 cyclic alkyl;
- b) -N(R20a)(R20b);
R20a and R20b are each independently hydrogen, substituted or unsubstituted C1-C12 linear, C3-C12 branched, or C3-C12 cyclic alkyl; substituted or unsubstituted C6 or C10 aryl; substituted or unsubstituted C1-C9 heterocyclic; substituted or unsubstituted C1-C11 heteroaryl; or R20a and R20b can be taken together to form a substituted or unsubstituted ring having from 3 to
10 carbon atoms and from 0 to 3 heteroatoms chosen from oxygen, nitrogen, and sulfur;
for example, -NHC(O)CH3, -CH2NHC(O)CH3, -NHC(O)NH2, -CH2NHC(O)NH2, -NHC(O)NHCH3, -CH2NHC(O)NHCH3, -OC(O)N(CH3)2, -CH2NHC(O)N(CH3)2, and the like;
xiii) -[C(R23a)(R23b)]xCN; for example; -CN, -CH2CN, and -CH2CH2CN;
xiv) -[C(R23a)(R23b)]xNO2; for example; NO2, -CH2NO2, and -CH2CH2NO2;
xv) -[C(R23a)(R23b)]xR21; for example, -CH2F, -CHF2, -CF3, -CCl3, or -CBr3;
R21 is C1-C10 linear, branched, or cyclic alkyl substituted by from 1 to 21 halogen atoms chosen
from -F, -Cl, -Br, or -I;
xvi) -[C(R23a)(R23b)]xSO2R22;
R22 is hydrogen, hydroxyl, substituted or unsubstituted C1-C4 linear or C3-C4 branched alkyl; substituted or unsubstituted C6, C10, or C14 aryl; C7-C15 alkylenearyl; substituted or unsubstituted C1-C9 heterocyclic; or substituted or unsubstituted C1-C11 heteroaryl; for example, -SO2H, -CH2SO2H, -SO2CH3, -CH2SO2CH3, - SO2C6H5, and -CH2SO2C6H5;
R
23a and R
23b are each independently hydrogen or C
1-C
4 alkyl; and
the index x is an integer from 0 to 5.
[0071] One aspect embodiment of this category relates to HIF-1α prolyl hydroxylase inhibitors
wherein R
1 and R
2 are taken together to form a 5-member substituted or unsubstituted C
1-C
4 heterocyclic or a substituted or unsubstituted C
1-C
4 heteroaryl ring, non-limiting examples of which include a ring chosen from:
i)
ii)
ii)
iii)
iv)
v)
vi)
vii)
or
viii)
[0072] Another aspect of this category relates to HIF-1α prolyl hydroxylase inhibitors wherein
R
1 and R
2 are taken together to form a substituted or unsubstituted C
4-C
11 heterocyclic or a substituted or unsubstituted C
4-C
11 heteroaryl ring, non-limiting examples of which are chosen from:
i)
ii)
iii)
iv)
or
v)
[0073] Non-limiting examples of this category include compounds having the formula:
and
[0074] A further category of the disclosed compounds has the formula:
wherein R represents from 1 to 5 optional substitutions for a phenyl ring hydrogen
atom, R
1 and R
2 are each independently hydrogen or substituted or unsubstituted C
1-C
10 linear or branched alkyl, wherein the alkyl unit can be substituted by one or more
units independently chosen from:
- i) C1-C8 linear, C3-C8 branched, or C3-C8 cyclic alkoxy;
- ii) hydroxy;
- iii) halogen;
- iv) cyano;
- v) amino, C1-C8 mono-alkylamino, C1-C8 di-alkylamino;
- vi) -SR40; R40 is hydrogen or C1-C4 linear or branched alkyl;
- vii) substituted or unsubstituted C6 of C10 aryl;
- viii) substituted or unsubstituted C1-C9 heterocyclic; or
- ix) substituted or unsubstituted C1-C9 heteroaryl.
[0075] Non-limiting examples of this category include:
[0076] A still yet further category of the disclosed HIF-1α prolyl hydroxylase inhibitors
relates to compounds having the formula:
wherein R
1 and R
2 are taken together to form a substituted or unsubstituted piperazine ring, the substitutions
on the ring as defined for R
200 herein above.
[0077] A yet still further category of the disclosed HIF-1α prolyl hydroxylase inhibitors
have the formula:
wherein R
1 and R
2 can be taken together to form a substituted or unsubstituted heterocyclic or heteroaryl
ring having from 2 to 20 carbon atoms and from 1 to 7 heteroatoms wherein the rings
formed exclude a a piperazine ring.
[0078] Also disclosed herein are
N-substituted benzyl or
N-substituted sulfonylaryl-3-hydroxypyridin-2-(1
H)-ones having the formula:
that can be used to stimulate the cellular immune response in a subject. For these
compounds, Z and L are the same as disclosed herein above. Non-limiting examples of
these compounds include:
1-(4-chlorobenzyl)-3-hydroxypyridin-2(1H)-one having the formula:
1-(-3-chlorobenzyl)-3-hydroxypyridin-2(1H)-one having the formula:
and
1-(2-chlorobenzyl)-3-hydroxypyridin-2(1H)-one having the formula:
[0079] Further disclosed herein are
N-substituted benzyl or
N-substituted sulfonylaryl-5-substituted-3-hydroxypyridin-2-(1
H)-ones having the formula:
wherein Y is substituted or unsubstituted phenyl, Z and L are the same as defined
herein above.
[0080] One aspect of Y relates to a phenyl group that is substituted with from 1 to 5 halogen
atoms, for example, Y is chosen from 2-chlorophenyl, 3-chlorophenyl, 2-fluorophenyl,
3-fluorophenyl, or 4-fluorophenyl. A further aspect of Y units relates to compounds
wherein Y is chosen from 2,3-difluorophenyl, 2,4-difluorophenyl, 2,5-difluorophenyl,
2,6-difluorophenyl, 2,3-dichlorophenyl, 2,4-dichlorophenyl, 2,5-dichlorophenyl, and
2,6-dichlorophenyl.
[0081] A non-limiting example of compounds according to this category include 1-(4-chlorobenzyl)-5-(4-chlorophenyl)-3-hydroxypyridin-2(1
H)-one having the formula:
[0082] Further non-limiting examples include:
1-(2-chlorobenzyl)-5-(2-chlorophenyl)-3-hydroxypyridin-2(1H)-one;
1-(2-chlorobenzyl)-5-(3-chlorophenyl)-3-hydroxypyridin-2(1H)-one;
1-(2-chlorobenzyl)-5-(4-chlorophenyl)-3-hydroxypyridin-2(1H)-one;
1-(3-chlorobenzyl)-5-(2-chlorophenyl)-3-hydroxypyridin-2(1H)-one;
1-(3-chlorobenzyl)-5-(3-chlorophenyl)-3-hydroxypyridin-2(1H)-one;
1-(3-chlorobenzyl)-5-(4-chlorophenyl)-3-hydroxypyridin-2(1H)-one;
1-(4-chlorobenzyl)-5-(2-chlorophenyl)-3-hydroxypyridin-2(1H)-one;
1-(4-chlorobenzyl)-5-(3-chlorophenyl)-3-hydroxypyridin-2(1H)-one;
1-(2-fluorobenzyl)-5-(2-chlorophenyl)-3-hydroxypyridin-2(1 H)-one;
1-(2-fluorobenzyl)-5-(3-chlorophenyl)-3-hydroxypyridin-2(1H)-one;
1-(2-fluorobenzyl)-5-(4-chlorophenyl)-3-hydroxypyridin-2(1H)-one;
1-(3-fluorobenzyl)-5-(2-chlorophenyl)-3-hydroxypyridin-2(1H)-one;
1-(3-fluorobenzyl)-5-(3-chlorophenyl)-3-hydroxypyridin-2(1H)-one;
1-(3-fluorobenzyl)-5-(4-chlorophenyl)-3-hydroxypyridin-2(1H)-one;
1-(4-fluorobenzyl)-5-(2-chlorophenyl)-3-hydroxypyridin-2(1H)-one;
1-(4-fluorobenzyl)-5-(3-chlorophenyl)-3-hydroxypyridin-2(1H)-one
1-(4-fluorobenzyl)-5-(4-chlorophenyl)-3-hydroxypyridin-2(1H)-one
1-(2-chlorobenzyl)-5-(2-fluorophenyl)-3-hydroxypyridin-2(1H)-one;
1-(2-chlorobenzyl)-5-(3-fluorophenyl)-3-hydroxypyridin-2(1H)-one;
1-(2-chlorobenzyl)-5-(4-fluorophenyl)-3-hydroxypyridin-2(1H)-one;
1-(3-chlorobenzyl)-5-(2-fluorophenyl)-3-hydroxypyridin-2(1H)-one;
1-(3-chlorobenzyl)-5-(3-fluorophenyl)-3-hydroxypyridin-2(1H)-one;
1-(3-chlorobenzyl)-5-(4-fluorophenyl)-3-hydroxypyridin-2(1H)-one;
1-(4-chlorobenzyl)-5-(2-fluorophenyl)-3-hydroxypyridin-2(1H)-one;
1-(4-chlorobenzyl)-5-(3-fluorophenyl)-3-hydroxypyridin-2(1H)-one;
1-(4-chlorobenzyl)-5-(3-fluorophenyl)-3-hydroxypyridin-2(1H)-one
1-(2-fluorobenzyl)-5-(2-fluorophenyl)-3-hydroxypyridin-2(1H)-one;
1-(2-fluorobenzyl)-5-(3-fluorophenyl)-3-hydroxypyridin-2(1H)-one;
1-(2-fluorobenzyl)-5-(4-fluorophenyl)-3-hydroxypyridin-2(1H)-one;
1-(3-fluorobenzyl)-5-(2-fluorophenyl)-3-hydroxypyridin-2(1H)-one;
1-(3-fluorobenzyl)-5-(3-fluorophenyl)-3-hydroxypyridin-2(1H)-one;
1-(3-fluorobenzyl)-5-(4-fluorophenyl)-3-hydroxypyridin-2(1H)-one;
1-(4-fluorobenzyl)-5-(2-fluorophenyl)-3-hydroxypyridin-2(1H)-one;
1-(4-fluorobenzyl)-5-(3-fluorophenyl)-3-hydroxypyridin-2(1H)-one; and
1-(4-fluorobenzyl)-5-(3-fluorophenyl)-3-hydroxypyridin-2(1H)-one.
[0083] The disclosed compounds are organized into several categories for the strictly non-limiting
purpose of describing alternatives for synthetic strategies for the preparation of
subgenera of compounds within the scope of the disclosed compounds that are not expressly
exemplified herein. This mental organization into categories does not imply anything
with respect to increased or decreased biological efficacy with respect to any of
the compounds or compositions of matter described herein.
[0084] Category I of the disclosed HIF-1α prolyl hydroxylase inhibitors relates to compounds
having the formula:
wherein A is a substituted or unsubstituted heterocyclic or heteroaryl ring having
from 2 to 20 carbon atoms and from 1 to 7 heteroatoms, R
200 represents from 0 to 40 substitutions form hydrogen, R represents from 1 to 5 substitutions
for hydrogen as defined herein above, and the index n is from 1 to 5. Table I provides
representative examples of compounds according to this category.
TABLE I
No. |
R |
A ring |
A1 |
3-methoxy |
pyrrolidin-1-yl |
A2 |
3-methoxy |
3-hydroxypyrrolidin-1-yl |
A3 |
3-methoxy |
2-(pyrdin-2-yl)pyrrolidin-1-yl |
A4 |
3-methoxy |
2-methylcarboxypyrrolidin-1-yl |
A5 |
3-methoxy |
2-(methoxymethyl)pyrrolidin-1-yl |
A6 |
3-methoxy |
thiazolidin-3-yl |
A7 |
3-methoxy |
1H-imidazol-1-yl |
A8 |
3-methoxy |
piperidin-1-yl |
A9 |
3-methoxy |
4-benzylpiperidin-1-yl |
A10 |
3-methoxy |
1,4'-bipiperidinyl-1'-yl |
A11 |
3-methoxy |
piperazin-1-yl |
A12 |
3-methoxy |
4-benzylpiperazin-1-yl |
A13 |
3-methoxy |
4-(2-methoxyphenyl)piperazin-1-ylmethyl |
A14 |
3-methoxy |
4-(6-chloropyridazin-3-yl)piperazin-1-yl |
A15 |
3-methoxy |
1,4-dioxa-8-azaspiro[4,5]dec-8-yl |
A16 |
3-methoxy |
morpholin-4-yl |
A17 |
3-methoxy |
thiomorpholin-4-yl |
A18 |
3-methoxy |
azepan-1-yl |
A19 |
3-methoxy |
azocan-1-vl |
A20 |
3-methoxy |
3,4-dihydroquinolin-1(2H)-yl |
A21 |
4-chloro |
pyrrolidin-1-yl |
A22 |
4-chloro |
3-hydroxypyrrolidin-1-yl |
A23 |
4-chloro |
2-(pyrdin-2-yl)pyrrolidin-1-yl |
A24 |
4-chloro |
2-methylcarboxypyrrolidin-1-yl |
A25 |
4-chloro |
2-(methoxymethyl)pyrrolidin-1-yl |
A26 |
4-chloro |
thiazolidin-3-yl |
A27 |
4-chloro |
1H-imidazol-1-yl |
A28 |
4-chloro |
piperidin-1-yl |
A29 |
4-chloro |
4-benzylpiperidin-1-yl |
A30 |
4-chloro |
1,4'-bipiperidinyl-1'-yl |
A31 |
4-chloro |
piperazin-1-yl |
A32 |
4-chloro |
4-benzylpiperazin-1-yl |
A33 |
4-chloro |
4-(2-methoxyphenyl)piperazin-1-ylmethyl |
A34 |
4-chloro |
4-(6-chloropyridazin-3-yl)piperazin-1-yl |
A35 |
4-chloro |
1,4-dioxB-8-azaspiro[4,5]dec-8-yl |
A36 |
4-chloro |
morpholin-4-yl |
A37 |
4-chloro |
thiomorpholin-4-yl |
A38 |
4-chloro |
azepan-1-yl |
A39 |
4-chloro |
azocan-1-yl |
A40 |
4-chloro |
3,4-dihydroquinolin-1(2H)-yl |
A41 |
4-chloro |
4-tert-butoxycarbonylpiperazin-1-yl |
[0085] The disclosed compounds of this category can be prepared by the procedure outlined
herein below in Scheme I and described in Example 1.
EXAMPLE 1
tert-Butyl-{[1-(4-chlorobenzyl)-3-hydroxy-2-oxo-1,2-dihydropyridin-4-yl]methyl}piperazine-1-carboxylate
(4)
[0086] Preparation of 3-(
tert-butyldimethylsilanyloxy)-1
H-pyridin-2-one (1): 3-Hydroxypyridin-2(1
H)-one (15 g, 135 mmol) and imidazole (23 g, 338 mmol) were suspended in dimethylformamide
(200 mL) under inert atmosphere. A solution of
tert-butyldimethylsilyl chloride (20.5 g, 136 mmol) in dimethylformamide (200 mL) is added
dropwise at room temperature over 30 minutes. The reaction was then allowed to stir
overnight. The resulting solution was then poured into water (300 mL) and the mixture
extracted with
tert-butyl methyl ether (3 x 500 mL). The combined organic layer was washed with water
(300 mL), brine (300 mL) then dried over Na
2SO
4. The solvent is removed under reduced pressure and the crude product crystallized
from heptanes to afford 16.3 g (53% yield) of the desired product.
1H NMR (250 MHz, CDCl
3) δ ppm 12.98 (1H, m); 6.91 (1H, dd, J = 1. Hz, J = 6.8 Hz); 6.81 (1H, dd, J = 1.8
Hz, J = 7.2 Hz); 6.02 - 6.007 (1H, m); 0.90 (9H, s), and 0.17 (6H, s).
[0087] Preparation of 3-(
tert-butyldimethylsilanyloxy)-1-(3-chlorobenzyl)-1
H-prydin-2-one (2): At 0 °C under an inert atmosphere, a solution of 4-chlorobenzyl
chloride (4.44 mmol) in THF (10 mL) was added dropwise to a solution of 3-(
tert-butyldimethylsilanyloxy)-1
H-pyridin-2-one, 1, (1 g, 4,44 mmol) and CsCO
3 (2.17 g, 6.66 mmol) in THF (10 mL). The reaction solution was allowed to warm to
room temperature and stirring was continued overnight. The resulting solution was
diluted with water (40 mL) and then extracted with EtOAc (3 x 30 mL). The combined
organic layer was washed with brine (30 mL) then dried over Na
2SO
4. The solvent is removed under reduced pressure and the crude product purified over
silica (EtOAc:heptane 4:1) to afford the desired product as a white solid.
[0088] Preparation of 1-(4-chlorobenzyl)-3-hydroxypyridin-2(1
H)-one (3): To a solution of 3-(
tert-butyldimethylsilanyloxy)-1-(3-chlorobenzyl)-1
H-prydin-2-one, 2, (2.36 g, 10 mmol) in EtOAc (25 mL) as added 5 M HCl (25 mL) with
vigorous stirring at room temperature. The reaction was monitored by TLC for the disappearance
of starting material and was complete within 30 minutes. The organic layer was decanted
and the aqueous phase extracted with dichloromethane (2 x 50 mL). The combined organic
layers were dried over Na
2SO
4 and the solvent removed under reduced pressure. The crude product was recrystallized
from dichloromethane. The yield was nearly quantitative.
1H NMR (360 MHz, DMSO-
d6) δ ppm 5.12 (2H, s); 6.13 (1 H, t,
J = 7.04); 6.71 (1H, dd,
J = 7.04, 1.59); 7.23-7.28 (2H, m); 7.36-7.43 (2H, m); 9.10 (1H, br. s).
[0089] Preparation of
tert-butyl-{[1-(4-chlorobenzyl)-3-hydroxy-2-oxo-1,2-dihydro-pyridin-4-yl]methyl}piperazine-1-carboxylate
(4):
tert-Butyl piperazine-1-carboxylate (97.6 mmol), formaldehyde (8 mL of a 37% soln., 97.6
mmol) and acetic acid (8 mL) were dissolved in ethanol (350 mL) and the solution stirred
for 1 hour at room temperature. A solution of 1-(4-chlorobenzyl)-3-hydroxypyridin-2(1
H)-one, 3, (48.8 mmol) in ethanol (350 mL) was added dropwise over 30 minutes. After
3 days of stirring, formaldehyde (3 mL) was added and the reaction heated to 50 °C
after which the reaction solution was concentrated under reduced pressure to approximately
500 mL. The desired product is obtained by crystallization from ethanol.
1H NMR (250 MHz, CDCl
3) d ppm 1.46 (s, 9H); 2.38-2.57 (m, 4H); 3.40-3.49 (m, 4H); 3.51 (s, 2H); 5.13 (s,
2H); 6.13 (d, J = 7.16 Hz), 1H); 6.79 (d, J = 7.16 Hz, 1H); 7.20-7.41 (m, 4H); 8.33-8.85
(m, 1H). The disclosed biological data relate to A41.
[0090] Category II of the disclosed prolyl hydroxylase inhibitors relates to compounds having
the formula:
wherein A is a substituted or unsubstituted heterocyclic or heteroaryl ring having
from 2 to 20 carbon atoms and from 1 to 7 heteroatoms, and R
200 represents from 0 to 40 substitutions form hydrogen. Table II provides representative
examples of compounds according to this category.
TABLE II
No. |
A ring |
B1 |
pyrrolidin-1-yl |
B2 |
3-hydroxypyrrolidin-1-yl |
B3 |
2-(pyrdin-2-yl)pyrrolidin-1-yl |
B4 |
2-methylcarboxypyrrolidin-1-yl |
B5 |
2-(methoxymethyl)pyrrolidin-1-yl |
B6 |
thiazolidin-3-yl |
B7 |
1H-imidazol-1-yl |
B8 |
piperidin-1-yl |
B9 |
4-benzylpiperidin-1-yl |
B10 |
1,4'-bipiperidinyl-1'-yl |
B11 |
piperazin-1-yl |
B12 |
4-benzylpiperazin-1-yl |
B13 |
4-(2-methoxyphenyl)piperazin-1-ylmethyl |
B14 |
4-(6-chloropyridazin-3-yl)piperazin-1-yl |
B15 |
1,4-dioxa-8-azaspiro[4,5]dec-8-yl |
B16 |
morpholin-4-yl |
B17 |
thiomorpholin-4-yl |
B18 |
azepan-1-yl |
B19 |
azocan-1-yl |
B20 |
3,4-dihydroquinolin-1(2H)-yl |
[0091] The compounds according to Category II can be prepared according to the procedure
outlined in Scheme I and disclosed in Example 1. The following are further examples
of inhibitors according to Category II.
[0092] 1-Benzyl-3-hydroxy-4-(piperidin-1-ylmethyl)pyridin-2(1H)-one: 1H NMR (300 MHz, CD
3OD) δ 1.81 (m, 6H), 3.07 (m, 2H), 3.51 (m, 2H), 4.23 (s, 2H), 5.24 (s, 2H), 6.31 (d,
J = 6.9 Hz, 1H), 7.35 (m, 6H);
19F NMR (252 MHz, CD
3OD) δ 85.5; 13C NMR (75 MHz, DMSO) δ 21.3, 22.7, 51.8, 52.5, 53.1, 106.4, 117.4, 127.7,
128.0, 128.2, 128.9, 137.3, 147.4, 158.0; ES MS(M+1) 299.12; HRMS Calcd. For C
18H
22N
2O
2, 298.38. Found (M+1)299.17.
[0093] 1-Benzyl-3-hydroxy-4-(morpholin-4-ylmethyl)pyridin-2(1H)-one: 1H NMR (300 MHz, DMSO) δ 3.25 (m, 4H), 3.81 (m, 4H), 4.18 (s, 2H), 5.17 (s, 2H), 6.31
(d,
J = 6.9 Hz, 1H), 7.35 (m, 6H);
19FNMR (300 MHz, DMSO) δ 88.5;
13C NMR (300 MHz, DMSO) δ 51.6, 51.8, 53.4, 63.5, 107.9, 119.1, 127.8, 128.0, 128.2,
128.9, 137.3, 147.5, 158.3; ES MS(M+1) 301.12; HRMS Calcd. For C
17H
20N
2O
3, 300.35.
[0094] 1-Benzyl-3-hydroxy-4-(thiomorpholin-4-ylmethyl)pyridin-2(1H)-one: 1HNMR(300 MHz, DMSO) δ 2.92 (m, 4H), 3.38 (m, 4H), 4.17 (s, 2H), 5.16 (s, 2H), 6.29
(d,
J = 7.5 Hz, 1H), 7.34 (m, 6H), 9.97 (s, 1H);
19F NMR (300 MHz, DMSO) δ 88.4;
13C NMR (75 MHz, DMSO) δ 24.3, 51.9, 53.4, 53.7, 107.9, 110.9, 127.8, 128.0, 128.2,
128.8, 137.2, 147.6, 157.6; ES MS (M+1) 317.14; HRMS Calcd. For C
17H
20N
2O
2S, 316.42. Found: (M+1) 317.13.
[0095] 1-Benzyl-3-hydroxy-4-(thiazolidin-3-ylmethyl)pyridin-2(1H)-one: 1HNMR (300 MHz, DMSO) δ 3.09 (t,
J = 6.3 Hz, 2H), 3.42 (t,
J = 6.3 Hz, 2H), 4.03 (s, 2H), 4.29 (s, 2H), 5.16 (s, 2H), 6.34 (d,
J= 7.2 Hz, 1H), 7.34 (m, 6H), 10.48 (broad s, 1H);
19FNMR (300 MHz, DMSO) δ 87.9;
13CNMR (75 MHz, DMSO) δ 28.3, 48.3, 50.1, 56.3, 57.0, 107.4, 122.1, 127.8, 128.2, 128.8,
137.4, 146.3, 157.6; ES MS (M+1) 303.08; Anal. Calcd for C
18H
19N
2O
4SF, C, 51.92; H, 4.60; N, 6.73; S, 7.70. Found: C, 51.67; H, 4.48; N, 6.69; S, 7.65.
[0096] 1-Benzyl-3-hydroxy-4-(pyrrolidin-1-ylmethyl)pyridin-2(1H)-one: 1H NMR (300 MHz, DMSO) δ 1.96 (s, 4H), 3.16 (s, 2H), 3.43 (s, 2H),4.23 (s, 4H), 5.17 (s, 2H),
6.34 (d,
J = 7.2 Hz, 1H), 7.34 (m, 6H);
19F NMR (252 MHz, DMSO) δ 88.7;
13C NMR (75 MHz, DMSO) δ 22.8, 50.9, 51.8, 53.7, 107.3, 118.0, 128.0, 128.2, 128.9,
137.3, 146.7, 157.6; ES MS (M+1) 285.13; Anal. Calcd. For C
19H
21F
3N
2O
4, C, 57.28; H, 5.31; N, 7.03. Found: C, 57.10; H, 5.11, N, 7.02.
[0097] 1-Benzyl-3-hydroxy-4-(4-benzylpiperidin-1-ylmethyl)pyridin-2(1H)-one: 1H NMR (DMSO) δ 1.43 (m, 2H), 1.72 (m, 4H), 2.96 (m, 2H), 3.41 (m, 3H), 4.09 (s, 2H),
5.16 (s, 2H), 6.34 (d,
J = 7.2 Hz, 1H), 7.35 (m, 11H);
19F NMR (252 MHz, DMSO) δ 88.8;
13C NMR (75 MHz, DMSO) δ; ES MS(M+1) 389.21; HRMS Calcd. For C
25H
28N
2O
2, 388.50. Found (M+1) 389.22.
[0098] 1-Benzyl-3-hydroxy-4-(4-benzylpiperazin-1-ylmethyl)pyridin-2(1H)-one: 1H NMR (300 MHz, DMSO) δ 3.11 (broad s, 4H), 3.81 (s, 2H), 4.18 (s, 2H), 5.15 (s, 2H),
6.24 (d,
J = 7.2 Hz, 1H), 7.34 (m, 6H), 7.46 (m, 5H);
19F NMR (252 MHz, DMSO) δ 88.2; 13C (75 MHz, DMSO) δ ; ES MS(M+1) 390.21; HRMS Calcd.
For C
24H
27N
3O
2, 389.49. Found (M+1) 390.21.
[0099] 1-Benzyl-3-hydroxy-4-[(3-hydroxypyrrolidin-1-yl)methyl]pyridin-2(1H)-one: 1HNMR (300 MHz, DMSO) δ 1.90 (m, 1H), 3.18 (m, 2H), 3.47 (m, 3H), 4.24 (s, 2H), 4.43
(s, 1H), 5.17 (s, 2H), 6.34 (d,
J = 7.2 Hz, 1H), 7.34 (m, 6H);
19F NMR (252 MHz, DMSO) δ 89.0;
13C NMR (75 MHz, DMSO) δ 51.8, 52.6, 61.3, 68.6, 107.4, 117.9, 128.0, 128.2, 128.9,
137.3, 146.7, 157.6; ES MS(M+1) 301.13; HRMS Calcd. For C
17H
20N
2O
3, 300.35. Found: (M+1) 301.15.
[0100] 1-Benzyl-3-hydroxy-4-(1,4-dioxa-8-azaspiro[4,5]dec-8-ylmethyl)pyridin-2(1H)-one: 1H NMR (300 MHz, DMSO) δ 1.90 (m, 4H), 3.11 (m, 2H), 3.43 (m, 2H), 3.93 (s, 4H), 4.19
(s, 2H), 5.16 (s, 2H), 6.34 (d,
J= 7.2 Hz, 1H), 7.34 (m, 6H), 10.01 (broad s, 1H);
19F NMR (252 MHz, DMSO) δ 88.3;
13C NMR (75 MHz, DMSO) δ 31.7, 50.7, 51.9, 52.5, 64.5, 101.1, 108.0, 116.5, 127.8, 128.0,
128.3, 128.9, 137.3, 147.5 157.6; ES MS(M+1) 357.19; HRMS Calcd. For C
20H
24N
4O
2, 356.42. Found(M+1) 357.18.
[0101] 1-Benzyl-3-hydroxy-4-azepan-1-ylmethylpyridin-2(1H)-one: 1H NMR (300 MHz, DMSO) δ 1.61 (m, 4H), 1.80 (m, 4H), 3.20 (m, 4H), 4.17 (s, 2H), 5.16
(s, 2H), 6.34 (d,
J = 7.2 Hz, 1H), 7.34 (m, 6H);
19F NMR (252 MHz, DMSO) δ 88.9;
13C NMR (75 MHz, DMSO) δ 22.8, 26.4, 51.8, 53.4, 54.4, 107.6, 117.2, 127.9, 128.0, 18.2,
128.9, 137.3, 147.2, 157.6; ES MS(M+1) 313.18; HRMS Calcd. For C
19H
24N
2O
4, 312.41. Found (M+1) 313.19.
[0102] 1-Benzyl-3-hydroxy-4-(azocan-1-ylmethyl)pyridin-2(1H)-one: 1H NMR (300 MHz, DMSO) δ 1.59 (m, 10H), 3.18 (m, 2H), 3.38 (m, 2H), 4.17 (s, 2H), 5.16
(s, 2H), 6.34 (d,
J = 7.2 Hz, 1H), 7.34 (m, 6H);
19F NMR (252 MHz, DMSO) δ 88.9;
13C NMR (75 MHz, DMSO) δ; ES MS(M+1) 327.2; HRMS Calcd. For C
20H
26N
2O
2, 326.43. Found (M+1) 327.20.
[0103] 1-Benzyl-3-hydroxy-(1,4'-bipiperidinyl-1'-ylmethyl)pyridin-2(1H)-one:
1H NMR (300 MHz, DMSO) δ 1.43-1.98 (m, 10H), 2.21 (m, 2H), 3.01 (m, 4H), 3.43 (m, 3H),
4.12 (s, 2H), 5.16 (s, 2H), 6.34 (d,
J = 7.2 Hz, 1H), 7.34 (m, 6H), 9.85 (broad s, 1H);
19F NMR (252 MHz, DMSO) δ 88.7; 1C NMR (75 MHz, DMSO) δ 21.6, 22.9, 23.8, 49.6, 50.5,
51.8, 53.0, 59.5, 108.0, 127.8, 128.0, 128.2, 128.9, 137.3, 147.5, 157.6; ES MS(M+1)
382.4; HRMS Calcd. For C
23H
31N
3O
2, 383.51. Found (M+1) 382.25.
[0104] 1-Benzyl-3-hydroxy-4-[(3,4-dihydroquinolin-1(2H)-yl)methyl]pyridin-2(1H)-one:
1H NMR (300 MHz, DMSO) δ 3.13 (t,
J = 6.3 Hz, 2H), 3.52 (m, 2H), 4.28 (s, 2H), 4.41 (s, 2H), 5.18 (s, 2H), 6.34 (d,
J = 7.2 Hz, 1H), 7.23-7.41 (m, 10H), 10.15 (broad s, 1H);
19F NMR (252 MHz, DMSO) δ 88.9;
13C NMR (75 MHz, DMSO) δ 25.4; 49.3, 51.8, 52.7, 52.9, 107.6, 11.6, 116.8, 126.9, 127.0,
127.9, 128.0, 128.1, 128.2, 128.8, 128.9, 131.7, 137.3, 147.3, 157.6; ES MS(M+1) 347.40;
HRMS Calcd. For C
22H
22N
2O
2, 346.42. Found (M+1)347.17.
[0105] Methyl 1-[(1-benzyl-3-hydroxy-2-oxo-1,2-dihydropyridin-4-yl)methyl]pyrrolidine-2-carboxylate: 1H NMR (300 MHz,.DMSO) δ 2.01 (m, 3H), 2.45 (m, 1H), 3.26 (m, 1H), 3.53 (m, 1H), 3.69
(s, 3H), 4.30 (m, 3H), 5.17 (s, 2H), 6.27 (d, 6.9 Hz, 1H), 7.35 (m, 6H),
19F NMR (252 MHz, DMSO) δ 88.3; 13C NMR (75 MHz, DMSO) δ; ES MS(M+1) 343.20; HRMS Calcd.
For C
19H
22N
2O
4, 342.39. Found (M+1)
[0106] 1-Benzyl-3-hydroxy-4-{[2-(methoxymethyl)pyrrolidin-1-yl]methyl}pyridin-2(1H)-one: 1H NMR (300 MHz, DMSO) δ 1.71 (m, 1H), 1.84 (m, 1H), 1.99 (m, 1H), 2.15 (m, 1H), 3.19
(m, 1H), 3.30 (s, 3H), 3.41 (m, 1H), 3.62 (m, 2H), 3.77 (m, 1H), 4.15 (m, 1H), 4.39
(m, 1H), 5.17 (s, 2H), 6.34 (d,
J = 7.2 Hz, 1H), 7.34 (m, 6H); 9.60 (broad s, 1H);
19F NMR (252 MHz, DMSO) δ 88.3;
13C NMR (75 MHz, DMSO) δ ; ES MS(M+1) 329.2; HRMS Calcd. For C
19H
24N
2O
3, 328.41. Found (M+1)
[0107] 1-Benzyl-3-hydroxy-4-{[2-(pyrdin-2-yl)pyrrolidin-1-yl]methyl}pyridin-2(1H)-one: 1H NMR (300 MHz, DMSO) δ 2.12 (m, 4H), 3.39 (m, 1H), 3.63 (m, 1H), 4.07 (m, 2H), 4.60
(m,. 1H), 5.10 (m, 2H), 6.15 (d,
J= 6.9 Hz, 1H), 7.33 (m, 6H), 7.44 (m, 1H), 8.05 (d,
J= 8.1 Hz, 1H), 8.59 (d,
J = 4.8 Hz, 1H), 8.74 (s, 1H);
19F NMR (252 MHz, DMSO) δ 88.0; ES MS(M+1) 362.22; HRMS Calcd. For C
22H
23N
3O
2, 361.44. Found (M+1).
[0108] 1-Benzyl-3-hydroxy-4-[4-(6-chloropyridazin-3-yl)piperazin-1-ylmethyl]pyridin-2(1H)-one: 1H NMR (300 MHz, DMSO) δ 3.18 (m, 2H), 3.48 (m, 4H), 4.19 (s, 2H), 4.46 (m, 2H), 5.16
(s, 2H), 6.62 (d,
J= 7.2 Hz, 1H), 7.35 (m, 6H), 7.48 (m, 1H), 7.68 (m, 1H), 11.5 (broad s, 1H);
13C NMR (75 MHz, DMSO) δ 42.1, 50.3, 51.9, 52.5, 108.2, 116.2; 118.0, 128.0, 128.2,
128.9, 129.8, 137.3, 147.4,. 157.6, 158.8; ES MS(M+1) 476.09. HRMS Calcd. For C
21H
22ClN
5N
3O
2, 411.88. Found (M+1) 412.76.
[0109] 1-Benzyl-3-hydroxy-4-[4-(2-methoxyphenyl)piperazin-1-ylmethyl]pyridin-2(1H)-one: 1H NMR (300 MHz, DMSO) δ 2.95 (m, 2H), 3.30 (m, 2H), 3.48 (m, 4H), 3.80 (s, 3H), 4.25
(s, 2H), 5.18 (s, 2H), 6.34 (d,
J = 7.2 Hz, 1H), 6.93 (m, 2H), 7.01 (m, 2H), 7.34 (m, 6H);
19F NMR (252 MHz, DMSO) δ 88.5; 13C NMR (75 MHz, DMSO) δ 47.2, 51.8, 53.0, 55.3, 108.1,
112.2, 114.8, 116.2, 118.6, 121.2, 123.8, 127.8, 128.0, 128.9, 137.3, 139.6, 147.5,
152.2, 157.6; ES MS(M+1) 405.82; HRMS Calcd. For C
24H
27N
3O
3, 405.49. Found (M+1) 406.21.
[0110] Category III of the disclosed prolyl hydroxylase inhibitors relates to compounds
having the formula:
[0111] R
1 and R
2 are each independently hydrogen or substituted or unsubstituted C
1-C
10 linear or branched alkyl, wherein the alkyl unit can be substituted by one or more
units independently chosen from:
- i) C1-C8 linear, C3-C8 branched, or C3-C8 cyclic alkoxy;
- ii) hydroxy;
- iii) halogen;
- iv) cyano;
- v) amino, C1-C8 mono-alkylamino, C1-C8 di-alkylamino;
- vi) -SR40; R40 is hydrogen or C1-C4 linear or C3-C4 branched alkyl;
- vii) substituted or unsubstituted C6 of C10 aryl;
- viii) substituted or unsubstituted C1-C9 heterocyclic; or
- ix) substituted or unsubstituted C1-C9 heteroaryl.
[0112] Table III herein below provides non-limiting examples of compounds encompassed by
this category.
TABLE III
No. |
R1 |
R2 |
C1 |
benzyl |
hydrogen |
C2 |
4-methoxybenzyl |
hydrogen |
C3 |
4-fluorobenzyl |
hydrogen |
C4 |
4-chlorobenzyl |
hydrogen |
C5 |
4-methylbenzyl |
hydrogen |
C6 |
2-(pyridin-2-yl)ethyl |
hydrogen |
C7 |
[1,3]dioxolan-2-ylmethyl |
hydrogen |
C8 |
tetrahydrofuran-2-ylmethyl |
hydrogen |
C9 |
2-methoxyethyl |
hydrogen |
C10 |
1-hydroxy-2-methylpropan-2-yl |
hydrogen |
C11 |
pyridin-4-ylmethyl |
hydrogen |
C12 |
furan-2-ylmethyl |
hydrogen |
C13 |
2-(methylthio)ethyl |
hydrogen |
C14 |
1-phenylethyl |
hydrogen |
C15 |
3-imidazol-1-ylpropyl |
hydrogen |
C16 |
cycloheptyl |
hydrogen |
C17 |
4-methylcyclohexyl |
hydrogen |
C18 |
1-benzylpiperidin-4-yl |
hydrogen |
C19 |
azepan-2-on-3-yl |
hydrogen |
C20 |
1-benzylpyrrolidin-3-yl |
hydrogen |
C21 |
benzyl |
methyl |
C22 |
4-methoxybenzyl |
methyl |
C23 |
4-fluorobenzyl |
methyl |
C24 |
4-chlorobenzvl |
methyl |
C25 |
4-methylbenzyl |
methyl |
C26 |
2-(pyridin-2-yl)ethyl |
methyl |
C27 |
[1,3]dioxolan-2-ylmethyl |
methyl |
C28 |
tetrahydrofuran-2-ylmethyl |
methyl |
C29 |
2-methoxyethyl |
methyl |
C30 |
1-hydroxy-2-methylpropan-2-yl |
methyl |
C31 |
pyridin-4-ylmethyl |
methyl |
C32 |
furan-2-ylmethyl |
methyl |
C33 |
2-(methylthio)ethyl |
methyl |
C34 |
1-phenylethyl |
methyl |
C35 |
3-(1H-imidazol-1-yl)propyl |
methyl |
C36 |
cycloheptyl |
methyl |
C37 |
4-methylcyclohexyl |
methyl |
C38 |
1-benzylpiperidin-4-yl |
methyl |
C39 |
azepan-2-on-3-yl |
methyl |
C40 |
1-benzylpyrrolidin-3-yl |
methyl |
[0113] The disclosed compounds of this category can be prepared by the procedure outlined
herein below in Scheme II and described in Example 2.
EXAMPLE 2
1-Benzyl-3-hydroxy-4-{[3-(1-H-imidazol-1-yl)propylamino]methyl}-pyridin-2(1H)-one (6)
[0114] N-Benzyl-3-hydroxypyridin-2(1
H)-one (5) can be prepared according to Example 1 by substituting benzyl bromide or
benzyl chloride into step (b) for (4-chloro)benzyl chloride.
[0115] 1-Benzyl-3-hydroxy-4-{[3-(1-
H-imidazol-1-yl)propylamino]methyl}pyridin-2(1
H)-one (6):
N-Benzyl-3-hydroxypyridin-2(1
H)-one (5) (250 mg, 1.23 mmol) and formaldehyde (200 mg, 273 eq.) are combined in aqueous
ethanol (10 mL) and stirred for 30 minutes. 3-(1-
H-Imidazol-1-yl)propan-1-amine (340 mg, 2.7 mmol) is then added and the reaction stirred
for 12 hours. The solvent is removed by evaporation and the residue dissolved in methanol
(2 mL) and purified via prep HPLC eluting with water/acetonitrile to afford the desired
product as the trifluoroacetate salt.
1H NMR (300 MHz, DMSO) δ 2.19 (m, 2H), 2.97 (m, 2H), 4.02 (s, 2H), 4.30 (t,
J = 6.6 Hz, 2H); 5.17 (s, 2H), 6.30 (d,
J = 6.9 Hz, 1H), 7.36 (m, 6H), 7.26 (s, 1H), 7.76 (s, 1H), 9.03 (s, 1H), 9.11 (s, 1H);
19F NMR (252 MHz, DMSO) δ 88.5;
13C NMR (75 MHz, DMSO) δ 6.5, 44.0, 46.0, 51.8, 106.8, 118.7, 120.5, 122.2, 127.9, 128.2,
128.9, 135.8, 137.4, 146.0, 158.2; ES MS(M+1) 339.05; HRMS Calcd. For C
19H
22N
4O
2, 338.44. Found (M+1) 339.18.
[0116] The following are further non-limiting examples of this aspect of the disclosed HIF-1α
prolyl hydroxylase inhibitors.
[0117] 1-Benzyl-3-hydroxy-4-(benzylaminomethyl)pyridin-2(1H)-one: 1HNMR (300 MHz, DMSO) δ 4.01 (s, 2H), 4.20 (s, 2H), 5.16 (s, 2H), 6.34 (d,
J = 7.2 Hz, 1H), 7.36 (m, 11H), 9.16 (broad s, 1H);
19FNMR(252 MHz, DMSO) δ 88.6;
13C NMR (75 MHz, DMSO) δ; ES MS(M+1) 321.16; Anal. Calcd. For C
22H
21F
3N
2O
4, C, 60.83; H, 4.87; N, 6.45. Found: C, 60.75; H, 4.56; N, 6.34.
[0118] 1-Benzyl-3-hydroxy-4-{[(2-(pyridin-2-yl)ethylamino]methyl}pyridin-2(1H)-one: 1H NMR (300 MHz, DMSO) δ 3.26 (m, 2H), 3.37 (m, 2H), 4.08 (s, 2H), 5.17 (s, 2H); 6.34
(d,
J = 7.2 Hz, 1H), 7.38 (m, 6H), 7.86 (d,
J = 5.7 Hz, 2H), 8.84 (m, 2H), 9.32 (broad s, 1H);
19FNMR(252 MHz, DMSO) δ 88.6;
13C NMR (75 MHz, DMSO) δ 31.5, 44.1, 46.3, 51.8, 106.9, 114.8, 127.1, 128.1, 128.8,
137.4, 143.8, 146.1, 155.3, 157.5, 158.4; ES MS (M+1) 336.18; HRMS Calcd For C
20H
21N
3O
2, 335.40. Found: 336.16.
[0119] 1-Benzyl-3-hydroxy-4-{[(tetrahydrofuran-2-ylmethyl)amino]methyl}pyridin-2(1H)-one: 1H NMR (300 MHz, DMSO) δ 1.56 (m, 1H), 1.86 (m, 2H), 1.99 (m, 1H), 2.92 (m, 1H), 3.05
(m, 1H), 3.80 (m, 2H), 4.09 (m, 3H), 5.16 (s, 2H), 6.34 (d,
J = 7.2 Hz, 1H), 7.34 (m, 6H); 8.91 (broad s, 1H);
19F NMR (252 MHz, DMSO) δ 88.5;
13C NMR(75 MHz, DMSO) δ ; ES MS(M+1) 315.16; HRMS. Calcd. For C
18H
22N
2O
3, 314.38. Found (M+1) 315.16.
[0120] 1-Benzyl-3-hydroxy-4-[(2-methoxyethylamino)methyl]pyridin-2(1H)-one: 1H NMR (300 MHz, DMSO) δ 3.13 (broad s, 2H), 3.30 (s, 3H), 3.59 (t,
J = 5.4 Hz, 2H), 4.02 (s, 2H), 5.16 (s, 2H), 6.34 (d,
J = 7.2 Hz, 1H), 7.34 (m, 6H), 8.91 (broad s, 1H);
19F NMR (252 MHz, DMSO) δ 88.4;
13C NMR (252 MHz, DMSO) δ ; ES MS(M+1) 289.13; HRMS Calcd. For C
16H
20N
2O
3, 288.34. Found (M+1) 289.15.
[0121] 1-Benzyl-3-hydroxy-4-[(1-hydroxy-2-methylpropan-2-ylamino)methyl]pyridin-2(1H)-one: 1H NMR (300 MHz, DMSO) δ 1.27 (s, 6H), 3.49 (s, 2H), 3.95 (s, 2H), 5.17 (s, 2H), 6.34
(d,
J = 7.2 Hz, 1H), 7.34 (m, 6H), 8.47 (broad s, 2H), 9.94 (broad s, 1H);
19F NMR (252 MHz, DMSO) δ 88.7; 13C NMR (75 MHz, DMSO) δ ; ES MS(M+1) 303.19; HRMS Calcd.
For C
17H
22N
2O
3, 302.37. Found (M+1) 303.17.
[0122] 1-Benzyl-3-hydroxy-4-[(pyridin-4-ylmethylamino)methyl]pyridin-2(1H)-one: 1H NMR (300 MHz, DMSO) δ 4.07 (s, 2H), 4.32 (s, 2H), 5,16 (s, 2H), 6.34 (d,
J = 7.2 Hz, 1H), 7.34 (m, 6H); 7.62 (d,
J = 5.7 Hz, 2H), 8.71 (d,
J = 4.5 Hz, 2H);
19F NMR (252 MHz, DMSO) δ 88.0;
13C NMR (75 MHz, DMSO) δ; ES MS(M+1) 322.17; HRMS Calcd. For C
19H
19N
3O
2, 321.37. Found (M+1) 322.15.
[0123] 1-Benzyl-3-hydroxy 4-{[(furan-2-ylmethyl)amino]methyl}pyridin-2(1H)-one: 1H NMR (300 MHz, DMSO) δ 4.00 (s, 2H), 4.28 (s, 2H), 5.16 (s, 2H), 6.27 (d,
J = 6.9 Hz, 1H), 6.54 (m, 1H), 6.65 (m ,1H), 7.34 (m, 6H), 7.80 (m, 1H), 9.27 (broad
s, 1H);
19F NMR (252 MHz, DMSO) δ 88.3;
13C NMR (75 MHz, DMSO) δ ; ES MS(M+1) 323.15; HRMS Calcd. For C
18H
18N
2O
3, 310.35. Found (M+1)
[0124] 1-Benzyl-3-hydroxy-4-{[2-(methylthio)ethylamino]methyl}pyridin-2(1H)-one: 1H NMR (300 MHz, DMSO) δ 2.10 (s, 3H), 2.74 (t,
J = 6.9 Hz, 2H), 3.16 (t,
J = 8.1 Hz, 2H), 4.05 (s, 2H), 5.17 (s, 2H), 6.34 (d,
J = 7.2 Hz, 1H), 7.34 (m, 6H),
19F NMR (252 MHz, DMSO) δ 89.0; ES MS(M+1) 305.14, HRMS Calcd. For C
16H
20N
2O
2S, 304.41. Found (M+1)
[0125] 1-Benzyl-3-hydroxy-4-[(4-methoxybenzylamino)methyl]pyridin-2(1H)-one: 1H NMR (300 MHz, DMSO) δ 3.70 (s, 3H), 3.98 (s, 2H), 4.13 (s, 2H), 5.16 (s, 2H), 6.28
(d,
J = 7.5 Hz, 1H), 7.00 (d,
J = 9.0 Hz, 4H), 7.34 (m, 6H); 9.07 (broad s, 1H);
19F NMR (252 MHz, DMSO) δ 89.0; ES MS(M+1) 351.10; HRMS Calcd. For C
21H
22N
2O
3, 350.41. Found (M+1) 351.17.
[0126] 1-Benzyl-3-hydroxy-4-[(1-phenylethylamino)methyl]pyridin-2(1H)-one: 1H NMR (300 MHz, DMSO) δ 1.59 (d,
J = 7.2 Hz, 3H), 3.71-3.93 (m, 2H), 4.45 (m, 1H), 5.15 (s, 2H), 6.28 (d,
J = 7.5 Hz, 1H), 7.34 (m, 11H);
19F NMR (252 MHz, DMSO) δ 88.9;
13C NMR (75 MHz, DMSO) δ 19.6, 42.5, 51.7, 58.0, 106.8, 119.3, 128.0, 128.1, 128.2,
128.9, 129.3, 129.4, 137.3, 145.9, 158.3; ES MS(M+1) 335.13; HRMS Calcd. For C
21H
22N
2O
2, 334.41. Found (M+1) 335.17.
[0127] 1-Benzyl-3-hydroxy-4-(cycloheptylaminomethyl)pyridin-2(1H)-one: 1H NMR (300 MHz, DMSO) δ 1.55 (m, 10H), 2.03 (m, 2H), 3.18 (s, 1H), 3.99 (m, 2H), 5.17
(s, 2H), 6.32 (d,
J = 6.9 Hz, 1H), 7.35 (m, 6H), 8.65 (broad s, 2H), 9.98 (broad s, 1H);
19F NMR (252 MHz, DMSO) δ 88.6;
13C NMR (75 MHz, DMSO) δ 23.0,27.2,30.4,41.6,51.7,58.9, 107.0, 111.7, 127.9, 128.0,
128.2, 128.8, 137.4, 146.0, 157.5; ES MS(M+1) 327.13; HRMS Calcd. For C
20H
26N
2O
2, 326.43. Found (M+1) 327.20.
[0128] 1-Benzyl-3-hydroxy-4-[(4-methylcyclohexylamino)methyl]pyridin-2(1H)-one: 1H NMR (300 MHz, DMSO) δ 0.93 (d,
J = 6.9 Hz, 3H), 1.38 (m, 4H),1.74 (m, 4H), 2.05 (m, 1H), 3.10 (m, 1H), 4.01 (s, 2H),
5.17 (s, 2H), 6.31 (m, 1H), 7.34 (m, 6H), 8.05 (broad s, 2H), 9.98 (broad s, 1H);
19F NMR (252 MHz, DMSO) δ 88.9; ES MS(M+1) 327.14; HRMS Calcd. For C
20H
26N
2O
2, 326.43; Found (M+1) 372.20.
[0129] 1-Benzyl-3-hydroxy-4-[(1-benzylpiperidin-4-ylamino)methyl]pyridin-2(1H)-one:
1H NMR (300 MHz, DMSO) δ 1.77 (m, 2H), 2.31 (m, 2H), 2.98 (m, 2H), 3.30 (m, 3H), 3.46
(m, 2H), 4.03 (s, 2H), .29 (s, 2H), 5.16 (s, 2H), 6.30 (d,
J = 7.5 Hz, 1H), 7.34 (m, 6H), 7.49 (s, 5H), 9.12 (broad s, 1H), 10.05 (broad s, 1H);
19F NMR (252 MHz, DMSO) δ 88.8;
13C NMR (75 MHz, DMSO) δ 27.1, 43.4, 51.8, 52.1, 54.2, 54.7, 57.6, 106.9, 118.5, 128.0,
128.1, 128.8, 129.3, 129.8, 130.7, 131.3, 137.3, 146.2, 157.4; ES MS(M+1) 404.56;
HRMS Calcd. For C
25H
28N
3O
2, 403.52. Found (M+1) 404.23.
[0130] 3-[(1-Benzyl-3-hydroxy-2-oxo-1,2-dihydropyridin-4-yl)methylamino]azepan-2-one:
1H NMR (300 MHz, DMSO) δ 1.25 (m, 1H), 1.59 (m, 2H), 1.74 (m, 1H), 1.92 (m, 1H), 2.10
(m, 1H), 3.18 (m, 3H), 4.03 (s, 2H), 4.2 (m, 1H), 5.17 (s, 2H), 6.33 (d,
J = 7.5 Hz, 1H), 7.34 (m, 6H), 8.31 (t,
J = 5.4 Hz, 1H), 9.07 (broad s, 2H), 9.90 (broad s, 1H);
19F NMR (252 MHz, DMSO) δ 88.4;
13C NMR (75 MHz, DMSO) δ 27.0, 27.2, 28.4, 43.4, 51.7, 59.3, 107.1, 118.9, 127.8, 127.9,
128.1, 128.9, 137.4, 146.0, 157.5, 166.3; ES MS(M+1) 342.01; HRMS Calcd. For C
19H
23N
3O
3, 341.40. Found (M+1) 342.18.
[0131] 1-Benzyl-3-hydroxy-4-[(1-benzylpyrrolidin-3-ylamino)methyl]pyridin-2(1H)-one: 1H NMR (300 MHz, DMSO) δ 2.22 (m, 2H), 2.42 (m, 1H), 3.39 (m, 3H), 3.68 (m, 1H), 4.06
(s, 2H), 4.39 (s, 2H), 5.17 (s, 2H), 6.33 (d,
J = 7.5 Hz, 1H), 7.30-7.52 (m, 11H);
19F NMR (252 MHz, DMSO) δ 88.5;
13C NMR (75 MHz, DMSO) δ 27.1, 43.4, 51.8, 52.1, 54.2, 54.7, 57.5, 106.9, 118.5, 128.0,
128.8, 129.3, 129.8, 130.7, 131.3, 137.3, 146.2, 157.5; ES MS(M+1) 390.14; HRMS Calcd.
For C
24H
27N
3O
2, 389.49. Found (M+1) 390.21.
[0132] (R)-1-Benzyl-3-hydroxy-4-[(1-phenylethylamino)methyl]pyridin-2(1H)-one: 1H NMR (300 MHz, DMSO) δ 1.58 (d, J = 6.9 Hz, 3H), 3.74 (m, 2H), 4.44 (m, 1H), 5.14
(s, 2H), 6.23 (d,
J = 7.2 Hz, 1H), 7.35 (m, 6H);
19F NMR (252 MHz, DMSO) δ 89.4;
13C NMR (75 MHz, DMSO) δ 19.6, 42.6, 51.7, 58.0, 106.9, 18.7, 128.0, 128.1, 128.8, 129.3,
129.4, 137.2, 137.4, 145.9, 157.5; ES MS(M+1) 335.13; Anal. Calcd. For C
21H
22N
2O
2, 334.41. Found (M+1) 335.31.
[0133] 1-Benzyl-3-hydroxy-4-[([1,3]dioxolan-2-ylmethylmethylamino)methyl]pyridin-2(1H)-one: 1H NMR (300 MHz, DMSO) δ 2.81 (s, 3H), 3.35 (d,
J = 3.9 Hz, 2H), 3.89 (m, 2H), 4.01 (m, 2H), 4.21 (m, 2H), 5.17 (s, 2H); 5.27 (t,
J = 3.9 Hz, 1H), 6.34 (d,
J = 7.2 Hz, 1H), 7.35 (m, 6H);
19F NMR (252 MHz, DMSO) δ 88.5;
13C NMR (75 MHz, DMSO) δ ; ES MS(M+1) 331.18; HRMS Calcd. For C
18H
22N
2O
4, 330.38. Found (M+1) 331.16.
[0134] Category IV of the disclosed prolyl hydroxylase inhibitors relates to compounds having
the formula:
wherein A represents a ring optionally substituted by one or more R
200 units. Table IV provides non-limiting examples of this category.
TABLE IV
No. |
A ring |
D1 |
pyrrolidin-1-yl |
D2 |
3-hydroxypvrrolidin-1-yl |
D3 |
2-(pyrdin-2-yl)pyrrolidin-1-yl |
D4 |
2-methylcarboxypyrrolidin-1-yl |
D5 |
2-(methoxymethyl)pyrrolidin-1-yl |
D6 |
thiazolidin-3-yl |
D7 |
1H-imidazol-1-yl |
D8 |
piperidin-1-yl |
D9 |
4-benzylpiperidin-1-yl |
D10 |
1,4' -bipiperidinyl-1'-yl |
D11 |
piperazin-1-yl |
D12 |
4-benzylpiperazin-1-yl |
D13 |
4-(2-methoxvphenyl)piperazin-1-ylmethyl |
D14 |
4-(6-chloropyridazin-3-yl)piperazin-1-yl |
D15 |
1,4-dioxa-8-azaspiro[4,5]dec-8-yl |
D16 |
morpholin-4-yl |
D17 |
thiomorpholin-4-yl |
D18 |
azepan-1-yl |
D19 |
azocan-1-yl |
D20 |
3,4-dihydroquinolin-1(2H)-yl |
[0135] The disclosed compounds of this category can be prepared by the procedure outlined
herein below in Scheme III and described in Example 3.
EXAMPLE 3
1-(4'-Methylbenzenesulfonyl)-3-hydroxy-4-(pyrrolidin-1-ylmethyl)pyridin-2(1H)-one (8)
[0136] 1-(4'-Methylbenzenesulfonyl)-3-hydroxypyridin-2(1H)-one (7): To stirred solution of 3-[(
tert-butyldimethylsilyl)oxy]pyridin-2(1
H)-one (1) (4.66 g, 20.7 mmol) in dry THF (150 mL), maintained at -78 °C under a dry
nitrogen atmosphere is added n-butyl lithium (1.6 M solution in hexane, 21.0 mmol).
After 20 minutes, 4-methyl-benzenesulfonyl chloride (3.95 g, 20.7 mmol) is added as
a THF solution. The solution is allowed to warm to room temperature over one hour,
the water (10 mL) is added and the contents of the reaction vessel is extracted with
EtOAc (3x), washed with brine (1x), dried over Na
2SO
4 and concentrated. The combined organic layers are dried over Na
2SO
4 and concentrated. The residue is taken up in ethanol (10 mL) and treated with conc.
HCl (2 mL). The mixture is allowed to stir for 1 hour and the solvent is removed under
reduced pressure to afford the desired compound as a white solid.
1H NMR (300 MHz, DMSO) δ 2.43 (s, 3H), 6.14 (t, J = 6.9 Hz, 1H), 6.76 (dd, J = 7.65
Hz, 1.5 Hz, 1H), 7.18 (dd, J = 6.6 Hz, 1.8 Hz, 1H), 7.32 (d, J = 7.3 Hz, 2H), 7. 98
(d, J = 7.9 Hz, 2H).
[0137] 1-(4'-Methylbenzenesulfonyl)-3-hydroxy4-(pyrrolidin-1-ylmethyl)pyridin-2(1H)-one (8): 1-(4'-Methylbenzenesulfonyl)-3-hydroxypyridin-2(1
H)-one (7) (250 mg, 0.94 mmol) and formaldehyde (200 mg, 2.07 mmol) are combined in
aqueous ethanol (10 mL) and stirred for 30 minutes. Pyrrolidine (149 mg, 2.07 mmol)
is then added and the reaction stirred for 12 hours. The solvent is removed by evaporation
and the residue dissolved in methanol (5 mL) and purified via prep HPLC eluting with
water/acetonitrile to afford the desired product.
1H NMR (300 MHz, DMSO) δ 1.87 (m, 2H), 1.99 (m, 2H), 2.44 (s, 3H), 3.09 (m, 2H), 3.40
(m, 2H), 4.19 (s, 2H), 6.51 (d,
J = 7.5 Hz, 1H), 7.51 (d,
J = 8.4 Hz, 1H), 7.76 (d,
J = 7.5 Hz, 1H), 7.98 (d,
J = 8.1 Hz, 1H), 9.93 (broad s, 1H);
19F NMR (252 MHz, DMSO) δ 88.4; 13C NMR (75 MHz, DMSO) δ 21.5, 22.7, 50.5, 53.7, 108.7,
118.6, 119.4, 128.4, 129.7, 130.1, 133.1, 146.8, 147.7, 156.2; ES MS(M+1) 349.25;
HRMS Calcd. For C
17H
20N
2O
4S, 348.42. Found (M+1) 349.42.
[0138] The following are further non-limiting examples of prolyl hydroxylase inhibitors
according to this category.
[0139] 1-(4'-Methylbenzenesulfonyl)-3-hydroxy-4-thiazolidin-3-ylmethylpyridin-2(1H)-one: 1H NMR (300 MHz, DMSO) δ 2.43 (s, 3H), 2.94 (t,
J = 6.6 MHz, 2H), 3.18 (t,
J = 6.0 Hz, 2H), 3.66 (s, 2H), 4.12 (s, 2H), 6.51 (d,
J = 7.5 Hz, 1H), 7.51 (d,
J = 8.4 Hz, 1H), 7.76 (d,
J = 7.5 Hz, 1H), 7.98 (d,
J = 8.1 Hz, 1H),
19F NMR (252 MHz, DMSO) δ 87.9;
13C NMR (75 MHz, DMSO) δ 21.5, 21.9, 24.6, 25.8, 50.3, 51.6, 108.7, 118.6, 120.8, 129.7,
130.1, 133.1, 146.9, 148.1, 156.1, 158.4, 158.8; ES MS(M+1) 367.18; HRMS Calcd. For
C
16H
18N
2O
4S
2, 366.46. Found (M+1) 367.43.
[0140] 1-(4'-Methylbenzenesulfonyl)-3-hydroxy-4-azocan-lylmethylpyridin-2(1H)-one: 1H NMR (300 MHz, DMSO) δ 1.59 (m, 10H), 2.44 (s, 3H), 3.17 (m, 2H), 3.32 (m, 2H), 4.15
(s, 2H), 6.51 (d,
J = 7.5 Hz, 1H), 7.51 (d,
J = 8.4 Hz, 1H), 7.76 (d,
J = 7.5 Hz, 1H), 7.98 (d,
J = 8.1 Hz);
19F NMR (252 MHz, DMSO) δ 88.7;
13C NMR (75 MHz, DMSO) δ 21.5, 21.9, 23.7, 24.6, 25.8, 50.3, 51.6, 108.7, 118.9, 120.8,
129.8, 130.1, 133.1, 146.9, 148.2, 156.1; ES MS(M+1) 391.18; HRMS Calcd. For C
20H
26N
2O
4S, 390.18. Found (M+1) 391.23.
[0141] 1-(4'-Methylbenzenesulfonyl)-3-hydroxy-4-(4-phenylpiperazin-1-ylmethyl)-pyridin-2(1H)-one: 1H NMR (300 MHz,. DMSO) δ 2.43 (s, 3H), 3.13 (m, 8H), 3.43 (s, 2H), 6.47 (d,
J = 7.5 Hz, 1H), 6.78 (t,
J = 7.2 Hz, 1H), 7.21 9m, 2H), 7.50 (d,
J = 8.1 Hz, 2H), 7.67 (d,
J = 7.8 Hz, 1H), 7.97 (d,
J = 8.4 Hz, 2H);
13C NMR (75 MHz, DMSO) δ 21.5, 42.6, 45.6, 46.2, 50.8, 51.9, 109.6, 116.4, 116.8, 117.7,
120.6, 121.1, 129.5, 129.6, 129.8, 130.1, 133.2, 146.8, 149.5, 156.1; ES MS(M+1) 440.15;
HRMS Calcd. For C
23H
25N
3O
5S, 439.53. Found (M+1) 440.16.
[0142] 1-(4'-Methylbenzenesulfonyl)-3-hydroxy-4-[1,4']Bipiperidinyl-1'-ylmethylpyridin-2(1H)-one: 1H NMR (300 MHz, DMSO) δ 1.43 (m, 1h), 1.67 (m, 2H), 1.82 (m, 4H), 2.19 (m, 2H), 2.44
(s, 3H), 2.94 (m, 4H), 3.39 (m, 2H), 3.54 (m, 3H), 4.06 (s, 2H), 6.47 (d,
J = 8.1 Hz, 1H), 7.51 (d,
J = 8.1 Hz, 2H), 7.73 (d, 7.8 Hz, 1H), 7.99 (d,
J = 8.4 Hz, 2H);
19F NMR (252 MHz, DMSO) δ 88.7;
13C NMR (75 MHz, DMSO) δ 21.4, 22.9, 23.6, 48.4, 49.5, 59.4, 109.3, 114.8, 117.6, 120.5,
122.7, 129.7, 130.1, 133.1, 146.9, 148.6, 156.2; ES MS(M+1) 446.19; HRMS Calcd. For
C
23H
31N
3O
4S, 445.58. Found (M+1) 446.21.
[0143] 1-(4'-Methylbenzenesulfonyl)-3-hydroxy-4-[4-(6-chloropyridazin-3-yl)piperazin-1-ylmethyl]pyridin-2(1H)-one: 1H NMR (300 MHz, DMSO) δ 2.44 (s, 3H), 3.17 (m, 2H), 3.46 (m, 4H), 4.17 (s, 2H), 4.45
(m, 2H), 6.77 (d,
J = 7.8 Hz, 1H), 7.04 (m, 1H), 7.53 (m 2H), 7.68 (m, 2H), 7.98 (m, 2H), 11.3 (broad
s, 1H), ES MS(M+1) 476.92. HRMS Calcd. For C
21H
25ClN
5O
4S, 475.95. Found (M+1) 476.11.
[0144] Category V of HIF-1α prolyl hydroxylase inhibitors relates to compounds having the
formula:
[0145] R represents from 1 to 5 optional substitutions for a phenyl ring hydrogen atom,
R
1 and R
2 are each independently hydrogen or substituted or unsubstituted C
1-C
10 linear or branched alkyl, wherein the alkyl unit can be substituted by one or more
units independently chosen from:
- i) C1-C8 linear, C3-C8 branched, or C3-C8 cyclic alkoxy;
- ii) hydroxy;
- iii) halogen;
- iv) cyano;
- v) amino, C1-C8 mono-alkylamino, C1-C8 di-alkylamino;
- vi) -SR40; R40 is hydrogen or C1-C4 linear or C3-C4 branched alkyl;
- vii) substituted or unsubstituted C6 of C10 aryl;
- viii) substituted or unsubstituted C1-C9 heterocyclic; or
- ix) substituted or unsubstituted C1-C9 heteroaryl.
[0146] Table V provides non-limiting examples of this category of HIF-1α prolyl hydroxylase
inhibitors.
TABLE V
No. |
R |
R1 |
R2 |
E1 |
4-methyl |
benzyl |
hydrogen |
E2 |
4-methyl |
4-methoxybenzyl |
hydrogen |
E3 |
4-methyl |
4-fluorobenzyl |
hydrogen |
E4 |
4-methyl |
4-chlorobenzyl |
hydrogen |
E5 |
4-methyl |
4-methylbenzyl |
hydrogen |
E6 |
4-methyl |
2-(pyridin-2-yl)ethyl |
hydrogen |
E7 |
4-methyl |
[1,3]dioxolan-2-ylmethyl |
hydrogen |
E8 |
4-methyl |
tetrahydrofuran-2-ylmethyl |
hydrogen |
E9 |
4-methyl |
2-methoxyethyl |
hydrogen |
E10 |
4-methyl |
1-hydroxy-2-methylpropan-2-yl |
hydrogen |
E11 |
4-methyl |
pyridin-4-ylmethyl |
hydrogen |
E12 |
4-methyl |
furan-2-ylmethyl |
hydrogen |
E13 |
4-methyl |
2-(methylthio)ethyl |
hydrogen |
E14 |
4-methyl |
1-phenylethyl |
hydrogen |
E15 |
4-methyl |
3-imidazol-1-ylpropyl |
hydrogen |
E16 |
4-methyl |
cycloheptyl |
hydrogen |
E17 |
4-methyl |
4-methylcyclohexyl |
hydrogen |
E18 |
4-methyl |
1-benzvlpiperidin-4-yl |
hydrogen |
E19 |
4-methyl |
azepan-2-on-3-yl |
hydrogen |
E20 |
4-methyl |
1-benzylpyrrolidin-3-yl |
hydrogen |
E21 |
4-methyl |
benzyl |
methyl |
E22 |
4-methyl |
4-methoxybenzyl |
methyl |
E23 |
4-methyl |
4-fluorobenzyl |
methyl |
E24 |
4-methyl |
4-chlorobenzyl |
methyl |
E25 |
4-methyl |
4-methylbenzyl |
methyl |
E26 |
4-methyl |
2-(pyridin-2-yl)ethyl |
methyl |
E27 |
4-methyl |
[1,3]dioxolan-2-ylmethyl |
methyl |
E28 |
4-methyl |
tetrahydrofuran-2-ylmethyl |
methyl |
E29 |
4-methyl |
2-methoxyethyl |
methyl |
E30 |
4-methyl |
1-hydroxy-2-methylpropan-2-yl |
methyl |
E31 |
4-methyl |
pyridin-2-ylmethyl |
methyl |
E32 |
4-methyl |
furan-2-ylmethyl |
methyl |
E33 |
4-methyl |
carboxymethyl |
methyl |
E34 |
4-methyl |
2-(methylthio)ethyl |
methyl |
E35 |
4-methyl |
1-phenylethyl |
methyl |
E36 |
4-methyl |
3-imidazol-1-ylpropyl |
methyl |
E37 |
4-methyl |
cycloheptyl |
methyl |
E38 |
4-methyl |
4-methylcyclohexyl |
methyl |
E39 |
4-methyl |
1-benzylpiperidin-4-yl |
methyl |
E40 |
4-methyl |
azepan-2-on-3-yl |
methyl |
E41 |
4-methyl |
1-benzylpyrrolidin-3-yl |
methyl |
[0147] The disclosed compounds of this category can be prepared by the procedure outlined
herein below in Scheme N and described in Examples 4.
EXAMPLE 4
1-(4'-Methylbenzenesulfonyl)-3-hydroxy-4-[(benzylamino)methyl]-pyridin-2(1H)-one (9)
[0148] 1-(4'-Methylbenzenesulfonyl)-3-hydroxy-4-(benzylaminomethyl)pyridin-2(1H)-one (9): 1 -(4'-Methylbenzene;ulfonyl)-3-hydroxypyridin-2(1
H)-one (7) (250 mg, 0.94 mmol) and formaldehyde (200 mg, 2.07 mmol) are combined in
aqueous ethanol (10 mL) and stirred for 30 minutes. Benzylamine (229 mg, 2.07 mmol)
is then added and the reaction stirred for 12 hours. The solvent is removed by evaporation
and the residue dissolved in methanol (5 mL) and purified via prep HPLC eluting with
water/acetonitrile to afford the desired product as the trifluoracetate salt.
1H NMR (300 MHz, DMSO) d 2.44 (s, 3H), 3.96 (s, 2H), 4.16 (s, 2H), 6.69 (d,
J = 8.1 Hz), 7.40 (m, 7H), 7.52 (m, 1H), 7.73 (d,
J = 8.1 Hz, 1H), 7.97 (d,
J = 8.1 Hz, 1H), 9.71 (broad s, 2H), 10.44 (broad s, 1H); ES MS(M+1 ) 396.67; HRMS
Calcd. For C
20H
20N
2O
4S, 384.45. Found (M+1) 385.12.
[0149] The following is a further non-limiting example of this category of HIF-1α prolyl
hydroxylase inhibitors.
[0150] 1-(4'-Methylbenzenesulfonyl)-3-hydroxy-4-[(2-methoxyethylamino)methyl]-pyridin-2(1H)-one: 1H NMR (300 MHz, DMSO) δ 2.43 (s, 3H), 3.12 (m, 2H), 3.29 (s, 3H), 3.56 (t,
J = 5.1 Hz, 2H), 3.99 (s, 2H), 6.51 (d,
J = 7.5 Hz, 1H), 7.51 (d,
J = 8.4 Hz, 1H), 7.76 (d,
J = 7.5 Hz, 1H), 7.98 (d,
J = 8.1 Hz);
19F NMR (252 MHz, DMSO) δ 88.6;
13C NMR (75 MHz, DMSO) δ 21.5, 43.8, 46.2, 46.5, 58.5, 67.2, 106.7, 119.2, 120.2, 123.9,
128.4, 129.7, 130.1, 133.1, 146.8, 147.0, 156.0; ES MS(M+1) 353.12. HRMS Calcd. For
C
16H
20N
2O
5S, 352.41. Found (M+1) 353.11.
[0151] Category VI of HIF-1α prolyl hydroxylase inhibitors relates to compounds having the
formula:
wherein L is chosen from CH
2 or SO
2 and Z is substituted or unsubstituted phenyl. Non-limiting examples of inhibitors
according to this category are disclosed in Table VI below.
TABLE VI
No. |
L |
Z |
F1 |
CH2 |
2-chlorophenyl |
F2 |
CH2 |
3-chlorophenyl |
F3 |
CH2 |
4-chlorophenyl |
F4 |
CH2 |
2-fluorophenyl |
F5 |
CH2 |
3-fluorophenyl |
F6 |
CH2 |
4-fluorophenyl |
F7 |
CH2 |
2,3-dichlorophenyl |
F8 |
CH2 |
2,4-dichlorophenyl |
F9 |
CH2 |
2,5-dichlorophenyl |
F10 |
CH2 |
2,6-dichlorophenyl |
F11 |
CH2 |
3,4-dichlorophenyl |
F12 |
CH2 |
3,5-dichlorophenyl |
F13 |
CH2 |
2,3-difluorophenyl |
F14 |
CH2 |
2,4-difluorophenyl |
F15 |
CH2 |
2,5-difluorophenyl |
F16 |
CH2 |
2,6-difluorophenyl |
F17 |
CH2 |
3,4-difluorophenyl |
F18 |
CH2 |
3,5-difluorophenyl |
F19 |
CH2 |
2-cyanophenyl |
F20 |
CH2 |
3-cyanophenyl |
F21 |
CH2 |
4-cyanophenyl |
F22 |
SO2 |
2-chlorophenyl |
F23 |
SO2 |
3-chlorophenyl |
F24 |
SO2 |
4-chlorophenyl |
F25 |
SO7 |
2-fluorophenyl |
F26 |
SO7 |
3-fluorophenyl |
F27 |
SO2 |
4-fluorophenyl |
F28 |
SO2 |
2,3-dichlorophenyl |
F29 |
SO2 |
2,4-dichlorophenyl |
F30 |
SO2 |
2,5-dichlorophenyl |
F31 |
SO2 |
2,6-dichlorophenyl |
F32 |
SO2 |
3,4-dichlorophenyl |
F33 |
SO2 |
3,5-dichlorophenyl |
F34 |
SO2 |
2,3-difluorophenyl |
F35 |
SO2 |
2,4-difluorophenyl |
F36 |
SO2 |
2,5-difluorophenyl |
F37 |
SO2 |
2,6-difluorophenyl |
F38 |
SO2 |
3,4-difluorophenyl |
F39 |
SO2 |
3,5-difluorophenyl |
F40 |
SO2 |
2-cyanophenyl |
F41 |
SO2 |
3-cyanophenyl |
F42 |
SO2 |
4-cyanophenyl |
[0152] The compounds encompassed within this category can be prepared according to Scheme
I for Z equal to CH
2 and according to Scheme III for Z equal to SO
2.
Pharmacetically Acceptable Salts
[0153] The disclosed HIF-1α prolyl hydroxylase inhibitors can be in the form of a pharmaceutically
acceptable salt. Pharmaceutically acceptable salts can be used by the formulator to
provide a form of the disclosed inhibitor that is more compatible with the intended
delivery of the inhibitor to a subject or for compatiblility of formulation.
[0154] The following are examples of procedures for preparing the pharmaceutically acceptable
salt of the disclosed inhibitor,
tert-butyl-{[1-(4-chlorobenzyl)-3-hydroxy-2-oxo-1,2-dihydropyridin-4-yl]methyl} piperazine-1-carboxylate.
[0155] A suspension of
tert-butyl-{[1-(4-chlorobenzyl)-3-hydroxy-2-oxo-1,2-dihydropyridin-4-yl]methyl}piperazine-1-carboxylate
(242 mg, 0.56 mmol) in MeOH (15 mL) was heated at reflux untill a homogeneous solution
was obtained. Heating was stopped and 0.1N HCI (6.7 mL, 1.2 eq.) was added while still
hot and the solution was cooled to room temperature. The volatiles were evaporated
under reduced pressure and the amorphous residue was crystallized in acetone (5 mL).
The solid was collected by filtration.
[0156] A suspension of
tert-butyl-{[1-(4-chlorobenzyl)-3-hydroxy-2-oxo-1,2-dihydropyridin-4-yl]methyl}piperazine-1-carboxylate
(217 mg, 0.5 mmol) in MeOH (15 mL) was heated at reflux untill a homogeneous solution
was obtained. Heating was stopped and methanesulfonic acid (115.2 mg, 1.2 eq.) was
added while still hot and the solution was cooled to room temperature. The volatiles
were evaporated under reduced pressure and the amorphous residue was crystallized
in acetone (5 mL). The solid was collected by filtration.
[0157] Table VII herein below provides examples of pharmaceutically acceptable salts of
tert-butyl-{[1-(4-chlorobenzyl)-3-hydroxy-2-oxo-1,2-dihydropyridin-4-yl]methyl}-piperazine-1-carboxylate
formed from organic and inorganic acids. Start
TABLE VII
Acid |
Yield |
Purity* |
M.P. (°C) |
color |
Free base |
- |
99.3% |
183-184 |
pink |
HCl |
90% |
99.7% |
185-186 |
white |
H2SO4 |
93% |
99.7% |
175 (dec.) |
slightly pink |
p-toluenesulfonyl |
74% |
99.8% |
185-186 |
white |
methanesulfonyl |
79% |
99.9% |
155-157 |
white |
[0158] 1H NMR analysis was used to determine the form of the salt, for example, that the mesylate
salt formed herein above had the following formula:
1H NMR analysis was used to determine at which site of the molecule salt formation
was taking place. The chemical shifts for the protons on the methylene group bridging
the piperazine and the pyridinone rings shifted from 3.59 ppm in the free base to
4.31 ppm of the salt. In addition, the piperazine methylene groups adjacent to the
tertiary amine shifted from 2.50 ppm to approximately 3.60 ppm. The chemical shifts
for the remaining protons were largely unchanged. These data indicate that the tertiary
amine nitrogen of the piperazine ring is protonated in salt forms. In addition, integration
of the methyl protons of the methane sulfonyl unit relative to the core compound indicates
the presence of one equivalent of the acid.
[0159] The formulator can determine the solubility of the pharmaceutically acceptable salts
of the disclosed inhibitors by any method desirable. The following is a non-limiting
example of a procedure for evaluating the solubility of a salt of a disclosed inhibitor.
A suspension of
tert-butyl-{[1-(4-chlorobenzyl)-3-hydroxy-2-oxo-1,2-dihydropyridin-4-yl]methyl}-piperazine-1-carboxylate
methanesulfonate (26.6 mg) in distilled deionized water (3.0 mL) is sonicated for
20 min with water bath temperature under 25 °C. The suspension is filtered to remove
any insoluble salt. The clear filtrate solution (200 µL) is diluted with distilled
deionized water (800 µL) and subjected to HPLC analysis. The following are results
for the pharmaceutically acceptable salts outlined in Table VII above.
Salt |
Solubility (mg/mL) |
Purity* |
Free base |
~0.001 |
99.3% |
hydrochloride |
5.9 |
99.7% |
hydrogensulfonate |
13.2 |
99.7% |
p-toluenesulfonate |
2.3 |
99.8% |
methanesulfonate |
16.6 |
99% |
[0160] The following are non-limiting examples of other acids that can be used to form pharmaceutically
acceptable salts of the disclosed inhibitors: acetate, ctrate, maleate, succinate,
lactate, glycolate and tartrate.
Step (a) Preparation of an O-protected hydroaypyridin-2(1H)-one
[0162] Step (a) relates to the formation of an O-protected hydroxypyridin-2(1
H)-one having the formula:
[0163] W can be any protecting group. Non-limiting examples of protecting groups include
carbamates, for example,
tert-butoxycarbonyl and methoxycarbonyl, alkylsilanes, for example, trimethylsilyl and
tert-butyldimethylsilyl, and the like.
Step (b) Preparation of O-protected N-benzyl hydroxypyridin-2(1H)-one or O-protected N-sulfonylphenyl hydroxypyridin-2(1H)-one
[0164] Step (b) relates to the formation of an O-protected
N-benzyl hydroxypyridin-2(1
H)-one or O-protected
N-sulfonylphenyl hydroxypyridin-2(1
H)-one having the formula
[0165] The protected hydroxypyridin-2(1
H)-one formed in step (a) is reacted with a compound having the formula:
wherein Q is a leaving group capable of being eliminated by the protected hydroxypyridin-2(1
H)-one ring nitrogen.
Step (c) Preparation of N-benzyl-3-hydroxypyridin-2(1H)-one or N-sulfonylphenyl-3-hydroxypyridin-2(1H)-one
[0166] Step (c) relates to the formation of an
N-benzyl-3-hydroxypyridin-2(1
H)-one or
N-sulfonylphenyl-3-hydroxypyridin-2(1
H)-one the having the formula:
[0167] Wherein the O-protected
N-benzyl hydroxypyridin-2(1
H)-one or O-protected
N-sulfonylphenyl hydroxypyridin-2(1
H)-one formed in step (b) is reacted with one or more reagents suitable for removing
protecting group W in a manner compatible with any R substitutions for hydrogen on
the phenyl ring.
Step (d) Preparation of an N-formylamine synthon
[0168] Step (d) relates to the formation of an
N-formylamine synthon having the formula:
[0169] The
N-formylamine is formed by reacting an amine having the formula:
with formaldehyde or a reagent capable of generating formaldehyde in situ.
Step (e) Preparation of the disclosed HIF-1α prolyl hydroxylase inhibitors
[0170] Step (e) relates to the formation of the final disclosed compounds having the formula:
by reacting the
N-formylamine formed in step (d) with the
N-benzyl-3-hydroxypyridin-2(11
H)-one or
N-sulfonylphenyl-3-hydroxypyridin-2(1
H)-one formed in step (c).
FORMULATIONS
Medicaments and Pharmaceutical Compositions
[0171] The present disclosure further relates to compositions or formulations that are useful
for making a medicament or a pharmaceutical composition. The disclosed medicaments
or pharmaceutical compositions comprising the disclosed human protein HIF-1α prolyl
hydroxylase inhibitors can comprise:
- a) an effective amount of one or more HIF-1α prolyl hydroxylase inhibitors according
to the present disclosure; and
- b) one or more excipients.
[0172] Diseases or conditions affected by increased stabilization of HIF-1 by inhibition
of HIF-1α prolyl hydroxylase include PVD, CAD, heart failure, ischemia, anemia, wound
healing, antimicrobial activity, increased phagocytosis, anti-cancer activity, and
increase in the effectiveness of vaccines.
[0173] For the purposes of the present disclosure the term "excipient" and "carrier" are
used interchangeably throughout the description of the present disclosure and said
terms are defined herein as, "ingredients which are used in the practice of formulating
a safe and effective pharmaceutical composition."
[0174] The formulator will understand that excipients are used primarily to serve in delivering
a safe, stable, and functional pharmaceutical, serving not only as part of the overall
vehicle for delivery but also as a means for achieving effective absorption by the
recipient of the active ingredient. An excipient may fill a role as simple and direct
as being an inert filler, or an excipient as used herein may be part of a pH stabilizing
system or coating to insure delivery of the ingredients safely to the stomach. The
formulator can also take advantage of the fact the compounds of the present disclosure
have improved cellular potency, pharmacokinetic properties, as well as improved oral
bioavailability.
[0175] Non-limiting examples of compositions according to the present disclosure include:
- a) from about 0.001 mg to about 1000 mg of one or more human protein HIF-1α prolyl
hydroxylase inhibitors according to the present disclosure; and
- b) one or more excipients.
[0176] Another example according to the present disclosure relates to the following compositions:
- a) from about 0.01 mg to about 100 mg of one or more human protein prolyl HIF-1α prolyl
hydroxylase inhibitors according to the present disclosure; and
- b) one or more excipients.
[0177] A further example according to the present disclosure relates to the following compositions:
- a) from about 0.1 mg to about 10 mg of one or more human protein HIF-1α prolyl hydroxylase
inhibitors according to the present disclosure; and
- b) one or more excipients.
[0178] A still further example of compositions according to the present disclosure comprise:
- a) an effective amount of one or more human protein HIF-1α prolyl hydroxylase inhibitors
according to the present disclosure; and
- b) one or more chemotherapeutic agents or chemotherapeutic compounds as further described
herein.
[0179] A yet still further example of compositions according to the present disclosure comprise:
- a) an effective amount of one or more human protein HIF-1a prolyl hydroxylase inhibitors
according to the present disclosure; and
- b) one or more vaccines for treatment of an infectious disease.
[0180] The present disclosure further relates to the use of one or more of the HIF-1α prolyl
hydroxylase inhibitors disclosed herein for making a medicament for treating anemia.
[0181] The present disclosure further relates to the use of one or more of the HIF-1α prolyl
hydroxylase inhibitors disclosed herein for making a medicament for treating increasing
cellular immunity.
[0182] The present disclosure further relates to the use of one or more of the HIF-1α prolyl
hydroxylase inhibitors disclosed herein for making a medicament for treating cancer.
[0183] The present disclosure further relates to the use of one or more of the HIF-1α prolyl
hydroxylase inhibitors disclosed herein for making a medicament for increasing HIF-1
stabilization.
[0184] The present disclosure further relates to the use of one or more of the HIF-1α prolyl
hydroxylase inhibitors disclosed herein for making a medicament for treating anemia.
[0185] The present disclosure further relates to the use of one or more of the HIF-1α prolyl
hydroxylase inhibitors disclosed herein for making a medicament for treating peripheral
vascular disease.
[0186] The present disclosure further relates to the use of one or more of the HIF-1α prolyl
hydroxylase inhibitors disclosed herein for making a medicament for treating wounds.
[0187] The present disclosure further relates to the use of one or more of the HIF-1α prolyl
hydroxylase inhibitors disclosed herein for making a medicament that is an antimicrobial.
[0188] The present disclosure further relates to the use of one or more of the HIF-1α prolyl
hydroxylase inhibitors disclosed herein for making a medicament for treating atherosclerotic
lesions.
[0189] The present disclosure further relates to the use of one or more of the HIF-1α prolyl
hydroxylase inhibitors disclosed herein for making a medicament for treating diabetes.
[0190] The present disclosure further relates to the use of one or more of the HIF-1α prolyl
hydroxylase inhibitors disclosed herein for making a medicament for treating hypertension.
[0191] The present disclosure further relates to the use of one or more of the HIF-1α prolyl
hydroxylase inhibitors disclosed herein for making a medicament for treating a disease
affected by the level of vascular endothelial growth factor (VEGF), glyceraldehyde
3-phosphate dehydrogenase (GAPDH), and erythropoietin (EPO).
[0192] The present disclosure further relates to the use of one or more of the HIF-1α prolyl
hydroxylase inhibitors disclosed herein for making a medicament for treating a disorder
chosen from Crohn's disease and ulcerative colitis, psoriasis, sarcoidosis, rheumatoid
arthritis, hemangiomas, Osler-Weber-Rendu disease, or hereditary hemorrhagic telangiectasia,
solid or blood borne tumors and acquired immune deficiency syndrome.
[0193] The present disclosure further relates to the use of one or more of the HIF-1α prolyl
hydroxylase inhibitors disclosed herein for making a medicament for treating a disorder
chosen from diabetic retinopathy, macular degeneration, cancer, sickle cell anemia,
sarcoid, syphilis, pseudoxanthoma elasticum, Paget's disease, vein occlusion, artery
occlusion, carotid obstructive disease, chronic uveitis/vitritis, mycobacterial infections,
Lyme's disease, systemic lupus erythematosis, retinopathy of prematurity, Eales' disease,
Behcet's disease, infections causing a retinitis or choroiditis, presumed ocular histoplasmosis,
Best's disease, myopia, optic pits, Stargardt's disease, pars planitis, chronic retinal
detachment, hyperviscosity syndrome, toxoplasmosis, trauma and post-laser complications,
diseases associated with rubeosis, and proliferative vitreoretinopathy.
[0194] The disclosed compositions and the form of pharmaceutical preparations comprising
the HIF-1α prolyl hydroxylase inhibitors alone, or in combination with another drug
or other therapeutic agent,
inter alia, chemotherapeutic agent or chemotherapeutic compound, can vary according to the intended
route of administration.
[0195] Orally administered preparations can be in the form of solids, liquids, emulsions,
suspensions, or gels, or in dosage unit form, for example as tablets or capsules.
Tablets can be compounded in combination with other ingredients customarily used,
such as tale, vegetable oils, polyols, gums, gelatin, starch, and other carriers.
The HIF-1α prolyl hydroxylase inhibitors can be dispersed in or combined with a suitable
liquid carrier in solutions, suspensions, or emulsions.
[0196] Parenteral compositions intended for injection, either subcutaneously, intramuscularly,
or intravenously, can be prepared as liquids or solid forms for solution in liquid
prior to injection, or as emulsions. Such preparations are sterile, and liquids to
be injected intravenously should be isotonic. Suitable excipients are, for example,
water, dextrose, saline, and glycerol.
[0197] Administration of pharmaceutically acceptable salts of the substances described herein
is included within the scope of the present disclosure. Such salts can be prepared
from pharmaceutically acceptable non-toxic bases including organic bases and inorganic
bases. Salts derived from inorganic bases include sodium, potassium, lithium, ammonium,
calcium, magnesium, and the like. Salts derived from pharmaceutically acceptable organic
non-toxic bases include salts of primary, secondary, and tertiary amines, basic amino
acids, and the like. For a helpful discussion of pharmaceutical salts, see
S.M. Berge et al., Journal of Pharmaceutical Sciences 66:1-19 (1977) the disclosure of which is hereby incorporated by reference.
[0198] Substances for injection can be prepared in unit dosage form in ampules, or in multidose
containers. The HIF-1α prolyl hydroxylase inhibitors or compositons comprising one
or more HIF-1α prolyl hydroxylase inhibitors to be delivered can be present in such
forms as suspensions, solutions, or emulsions in oily or preferably aqueous vehicles.
Alternatively, the salt of the HIF-1α prolyl hydroxylase inhibitor can be in lyophilized
form for reconstitution, at the time of delivery, with a suitable vehicle, such as
sterile pyrogen-free water. Both liquids as well as lyophilized forms that are to
be reconstituted will comprise agents, preferably buffers, in amounts necessary to
suitably adjust the pH of the injected solution. For any parenteral use, particularly
if the formulation is to be administered intravenously, the total concentration of
solutes should be controlled to make the preparation isotonic, hypotonic, or weakly
hypertonic. Nonionic materials, such as sugars, are preferred for adjusting tonicity,
and sucrose is particularly preferred. Any of these forms can further comprise suitable
formulatory agents, such as starch or sugar, glycerol or saline. The compositions
per unit dosage, whether liquid or solid, can contain from 0.1% to 99% of polynucleotide
material.
METHODS
Methods Relating to Stabilization of HIF-1
[0199] The eradication of invading microorganisms depends initially on innate immune mechanisms
that preexist in all individuals and act within minutes of infection. Phagocytic cell
types, including macrophages and neutrophils, play a key role in innate immunity because
they can recognize, ingest, and destroy many pathogens without the aid of an adaptive
immune response. The effectiveness of myeloid cells in innate defense reflects their
capacity to function in low oxygen environments. Whereas in healthy tissues oxygen
tension is generally 20-70 mm HG (i.e. 2.5-9% oxygen), much lower levels (<1% oxygen)
have been described in wounds and necrotic tissue foci (
Arnold et al., Br J Exp Pathol 68, 569 (1987);
Vogelberg & Konig, Clin Investig 71, 466 (1993);
Negus et al., Am J Pathol 150, 1723 (1997)). It has also been shown (
Zinkernagel A. S. et al., "Pharmacologic Augmentation of Hypoxia-Inducible Factor-1α
with Mimosine Boosts the Bactericidal Capacity of Phagocytes" J. Infectious Diseases
(2008):197: 214-217) that the HIF-1α agonist mimosine can boost the capacity of human phagocytes and
whole blood to kill the leading pathogen
Staphylococcus aureus in a dose-dependent fashion and reduce the lesion size in a murine model of
S. aureus skin infection.
[0200] Macrophages are one population of effector cells involved in immune responses. Their
role in natural immunity includes mediation of phagocytosis, as well as release of
cytokines and cytotoxic mediators. They also facilitate the development of acquired
immunity through antigen presentation and release of immunomodulatory cytokines. Although
macrophages are immune effectors, they are also susceptible to infection by agents
such as bacteria, protozoa, parasites, and viruses (
The Macrophage, C. E. Lewis & J.O'D. McGee. eds., IRL Press at Oxford University Press,
New York, N.Y., 1992). Viruses capable of infecting macrophages include several RNA viruses such as measles
virus (MV) (e.g.,
Joseph et al., J. Virol. 16, 1638-1649, 1975), respiratory syncytial virus (RSV) (
Midulla et al., Am. Rev. Respir. Dis. 140, 771-777, 1989), and human immunodeficiency virus type 1 (HIV-1) (
Meltzer and Gendelman, in Macrophage Biology and Activation, S. W. Russell and S.
Gordon, eds., Springer-Verlag, New York, N.Y., pp. 239-263(1992:
Potts et al., Virology 175, 465-476, 1990).
[0201] Disclosed herein is a method for increasing HIF-1 stabilization in a cell, comprising
contacting a cell
in vivo, in vitro, or
ex vivo with an effective amount of one or more of the disclosed HIF-1α prolyl hydroxylase
inhibitors.
[0202] Also disclosed herein are methods for increasing the cellular immune response of
a human or mammal in need of increased cellular immunity, comprising administering
to a human or mammal in need with an effective amount of one or more of the disclosed
HIF-1α prolyl hydroxylase inhibitors.
[0203] Further disclosed herein are methods for increasing the cellular immune response
of a human or mammal diagnosed with a medical condition causing a decreased cellular
immunity, comprising administering to a human or mammal in need with an effective
amount of one or more of the disclosed HIF-1α prolyl hydroxylase inhibitors.
[0204] Yet further disclosed herein are methods for increasing the cellular immune response
of a human or mammal diagnosed with a medical condition causing a decreased cellular
immunity, comprising administering to a human or mammal in need with an effective
amount of one or more of the disclosed HIF-1α prolyl hydroxylase inhibitors.
[0205] Still further disclosed herein are methods for increasing the cellular immune response
of a human or mammal having a medical condition causing a decreased cellular immunity,
comprising administering to a human or mammal in need with an effective amount of
one or more of the disclosed HIF-1α prolyl hydroxylase inhibitors.
[0206] As such, the one or more HIF-1α prolyl hydroxylase inhibitor and any co-administered
compounds can be administered or contacted with a cell topically, buccally, orally,
intradermally, subcutaneously, mucosally in the eye, vagina, rectum, and nose, intravenously,
and intramuscularly
Methods Relating to the Treatment of Cancer
[0207] As used herein cancer is defined herein as "an abnormal growth of cells which tend
to proliferate in an uncontrolled way and, in some cases, to metastasize." As such,
both metastatic and non-metastatic cancers can be treated by the disclosed methods.
[0208] Disclosed are methods for treating cancer in a human or mammal, comprising administering
to a human or mammal with a cancer with an effective amount of one or more of the
disclosed HIF-1α prolyl hydroxylase inhibitors.
[0209] Also disclosed herein are methods for treating a human or mammal diagnosed with cancer,
co-administering to a human or mammal one or more chemotherapeutic agent or chemotherapeutic
compound together with one or more of the disclosed HIF-1α prolyl hydroxylase inhibitors.
[0210] The following are non-limiting examples of malignant and non-malignant cancers. Acute
Lymphoblastic; Acute Myeloid Leukemia; Adrenocortical Carcinoma; Adrenocortical Carcinoma,
Childhood; Appendix Cancer; Basal Cell Carcinoma; Bile Duct Cancer, Extrahepatic;
Bladder Cancer; Bone Cancer; Osteosarcoma and Malignant Fibrous Histiocytoma; Brain
Stem Glioma, Childhood; Brain Tumor, Adult; Brain Tumor, Brain Stem Glioma, Childhood;
Brain Tumor, Central Nervous System Atypical Teratoid/Rhabdoid Tumor, Childhood; Central
Nervous System Embryonal Tumors; Cerebellar Astrocytoma; Cerebral Astrocytoma/Malignant
Glioma; Craniopharyngioma; Ependymoblastoma; Ependymoma; Medulloblastoma; Medulloepithelioma;
Pineal Parenchymal Tumors of Intermediate Differentiation; Supratentorial Primitive
Neuroectodermal Tumors and Pineoblastoma; Visual Pathway and Hypothalamic Glioma;
Brain and Spinal Cord Tumors; Breast Cancer; Bronchial Tumors; Burkitt Lymphoma; Carcinoid
Tumor; Carcinoid Tumor, Gastrointestinal; Central Nervous System Atypical Teratoid/Rhabdoid
Tumor; Central Nervous System Embryonal Tumors; Central Nervous System Lymphoma; Cerebellar
Astrocytoma; Cerebral Astrocytoma/Malignant Glioma, Childhood; Cervical Cancer; Chordoma,
Childhood; Chronic Lymphocytic Leukemia; Chronic Myelogenous Leukemia; Chronic Myeloproliferative
Disorders; Colon Cancer; Colorectal Cancer; Craniopharyngioma; Cutaneous T-Cell Lymphoma;
Esophageal Cancer; Ewing Family of Tumors; Extragonadal Germ Cell Tumor; Extrahepatic
Bile Duct Cancer; Eye Cancer, Intraocular Melanoma; Eye Cancer, Retinoblastoma; Gallbladder
Cancer; Gastric (Stomach) Cancer; Gastrointestinal Carcinoid Tumor; Gastrointestinal
Stromal Tumor (GIST); Germ Cell Tumor, Extracranial; Germ Cell Tumor, Extragonadal;
Germ Cell Tumor, Ovarian; Gestational Trophoblastic Tumor; Glioma; Glioma, Childhood
Brain Stem; Glioma, Childhood Cerebral Astrocytoma; Glioma, Childhood Visual Pathway
and Hypothalamic; Hairy Cell Leukemia; Head and Neck Cancer; ;Hepatocellular (Liver)
Cancer; Histiocytosis, Langerhans Cell; Hodgkin Lymphoma; Hypopharyngeal Cancer; Hypothalamic
and Visual Pathway Glioma; Intraocular Melanoma; Islet Cell Tumors; Kidney (Renal
Cell) Cancer; Langerhans Cell Histiocytosis; Laryngeal Cancer; Leukemia, Acute Lymphoblastic;
Leukemia, Acute Myeloid; Leukemia, Chronic Lymphocytic; Leukemia, Chronic Myelogenous;
Leukemia, Hairy Cell; Lip and Oral Cavity Cancer; Liver Cancer; Lung Cancer, Non-Small
Cell; Lung Cancer, Small Cell; Lymphoma, AIDS-Related; Lymphoma, Burkitt; Lymphoma,
Cutaneous T-Cell; Lymphoma, Hodgkin; Lymphoma, Non-Hodgkin; Lymphoma, Primary Central
Nervous System; Macroglobulinemia, Waldenström; Malignant Fibrous Histiocytoma of
Bone and Osteosarcoma; Medulloblastoma; Melanoma; Melanoma, Intraocular (Eye); Merkel
Cell Carcinoma; Mesothelioma; Metastatic Squamous Neck Cancer with Occult Primary;
Mouth Cancer; Multiple Endocrine Neoplasia Syndrome, (Childhood); Multiple Myeloma/Plasma
Cell Neoplasm; Mycosis Fungoides; Myelodysplastic Syndromes; Myelodysplastic/Myelo-proliferative
Diseases; Myelogenous Leukemia, Chronic; Myeloid Leukemia, Adult Acute; Myeloid Leukemia,
Childhood Acute; Myeloma, Multiple; Myeloproliferative Disorders, Chronic; Nasal Cavity
and Paranasal Sinus Cancer; Nasopharyngeal Cancer; Neuroblastoma; Non-Small Cell Lung
Cancer; Oral Cancer; Oral Cavity Cancer; Oropharyngeal Cancer; Osteosarcoma and Malignant
Fibrous Histiocytoma of Bone; Ovarian Cancer; Ovarian Epithelial Cancer; Ovarian Germ
Cell Tumor; Ovarian Low Malignant Potential Tumor; Pancreatic Cancer; Pancreatic Cancer,
Islet Cell Tumors; Papillomatosis; Parathyroid Cancer; Penile Cancer; Pharyngeal Cancer;
Pheochromocytoma; Pineal Parenchymal Tumors of Intermediate Differentiation; Pineoblastoma
and Supratentorial Primitive Neuroectodermal Tumors; Pituitary Tumor; Plasma Cell
Neoplasm/Multiple Myeloma; Pleuropulmonary Blastoma; Primary Central Nervous System
Lymphoma; Prostate Cancer; Rectal Cancer; Renal Cell (Kidney) Cancer; Renal Pelvis
and Ureter, Transitional Cell Cancer; Respiratory Tract Carcinoma Involving the
NUT Gene on Chromosome 15; Retinoblastoma; Rhabdomyosarcoma; Salivary Gland Cancer; Sarcoma,
Ewing Family of Tumors; Sarcoma, Kaposi; Sarcoma, Soft Tissue; Sarcoma, Uterine; Sézary
Syndrome; Skin Cancer (Nonmelanoma); Skin Cancer (Melanoma); Skin Carcinoma, Merkel
Cell; Small Cell Lung Cancer; Small Intestine Cancer; Soft Tissue Sarcoma; Squamous
Cell Carcinoma, Squamous Neck Cancer with Occult Primary, Metastatic; Stomach (Gastric)
Cancer; Supratentorial Primitive Neuroectodermal Tumors; T-Cell Lymphoma, Cutaneous;
Testicular Cancer; Throat Cancer; Thymoma and Thymic Carcinoma; Thyroid Cancer; Transitional
Cell Cancer of the Renal Pelvis and Ureter; Trophoblastic Tumor, Gestational; Urethral
Cancer; Uterine Cancer, Endometrial; Uterine Sarcoma; Vaginal Cancer; Vulvar Cancer;
Waldenström Macroglobulinemia; and Wilms Tumor
[0211] Further disclosed herein are methods for treating cancer in a human or mammal, comprising
co-administering to a human or mammal, together with one or more chemotherapeutic
agents or chemotherapeutic compounds, one or more of the disclosed HIF-1α prolyl hydroxylase
inhibitors.
[0212] Also disclosed herein are methods for treating a human or mammal diagnosed with cancer,
co-administering to a human or mammal, together with one or more chemotherapeutic
agent or chemotherapeutic compound one or more of the disclosed HIF-1α prolyl hydroxylase
inhibitors.
[0213] A "chemotherapeutic agent" or "chemotherapeutic compound" is a chemical compound
useful in the treatment of cancer. The chemotherapeutic cancer agents that can be
used in combination with the disclosed HIF-1α inhibitors include, but are not limited
to, mitotic inhibitors (vinca alkaloids). These include vincristine, vinblastine,
vindesine and Navelbine™ (vinorelbine,5'-noranhydroblastine). In yet other embodiments,
chemotherapeutic cancer agents include topoisomerase I inhibitors, such as camptothecin
compounds. As used herein, "camptothecin compounds" include Camptosar™ (irinotecan
HCL), Hycamtin™ (topotecan HCL) and other compounds derived from camptothecin and
its analogues. Another category of chemotherapeutic cancer agents that may be used
in the methods and compositions disclosed herein are podophyllotoxin derivatives,
such as etoposide, teniposide and mitopodozide. The present disclosure further encompasses
other chemotherapeutic cancer agents known as alkylating agents, which alkylate the
genetic material in tumor cells. These include without limitation cisplatin, cyclophosphamide,
nitrogen mustard, trimethylene thiophosphoramide, carmustine, busulfan, chlorambucil,
belustine, uracil mustard, chlomaphazin, and dacarbazine. The disclosure encompasses
antimetabolites as chemotherapeutic agents. Examples of these types of agents include
cytosine arabinoside, fluorouracil, methotrexate, mercaptopurine, azathioprime, and
procarbazine. An additional category of chemotherapeutic cancer agents that may be
used in the methods and compositions disclosed herein include antibiotics. Examples
include without limitation doxorubicin, bleomycin, dactinomycin, daunorubicin, mithramycin,
mitomycin, mytomycin C, and daunomycin. There are numerous liposomal formulations
commercially available for these compounds. The present disclosure further encompasses
other chemotherapeutic cancer agents including without limitation anti-tumor antibodies,
dacarbazine, azacytidine, amsacrine, melphalan, ifosfamide and mitoxantrone.
[0214] The disclosed HIF-1α prolyl hydroxylase inhibitors herein can be administered in
combination with other anti-tumor agents, including cytotoxic/antineoplastic agents
and anti-angiogenic agents. Cytotoxic/anti-neoplastic agents are defined as agents
which attack and kill cancer cells. Some cytotoxic/anti-neoplastic agents are alkylating
agents, which alkylate the genetic material in tumor cells, e.g., cis-platin, cyclophosphamide,
nitrogen mustard, trimethylene thiophosphoramide, carmustine, busulfan, chlorambucil,
belustine, uracil mustard, chlomaphazin, and dacabazine. Other cytotoxic/anti-neoplastic
agents are antimetabolites for tumor cells, e.g., cytosine arabinoside, fluorouracil,
methotrexate, mercaptopuirine, azathioprime, and procarbazine. Other cytotoxic/anti-neoplastic
agents are antibiotics, e.g., doxorubicin, bleomycin, dactinomycin, daunorubicin,
mithramycin, mitomycin, mytomycin C, and daunomycin. There are numerous liposomal
formulations commercially available for these compounds. Still other cytotoxic/anti-neoplastic
agents are mitotic inhibitors (vinca alkaloids). These include vincristine, vinblastine
and etoposide. Miscellaneous cytotoxic/anti-neoplastic agents include taxol and its
derivatives, L-asparaginase, anti-tumor antibodies, dacarbazine, azacytidine, amsacrine,
melphalan, VM-26, ifosfamide, mitoxantrone, and vindesine.
[0215] Anti-angiogenic agents are well known to those of skill in the art. Suitable anti-angiogenic
agents for use in the disclosed methods and compositions include anti-VEGF antibodies,
including humanized and chimeric antibodies, anti-VEGF aptamers and antisense oligonucleotides.
Other known inhibitors of angiogenesis include angiostatin, endostatin, interferons,
interleukin 1 (including α and β) interleukin 12, retinoic acid, and tissue inhibitors
of metalloproteinase-1 and -2. (TIMP-1 and -2). Small molecules, including topoisomerases
such as razoxane, a topoisomerase II inhibitor with anti-angiogenic activity, can
also be used.
[0216] Other anti-cancer agents that can be used in combination with the disclosed HIF-1α
inhibitors include, but are not limited to: acivicin; aclarubicin; acodazole hydrochloride;
acronine; adozelesin; aldesleukin; altretamine; ambomycin; ametantrone acetate; aminoglutethimide;
amsacrine; anastrozole; anthramycin; asparaginase; asperlin; azacitidine; azetepa;
azotomycin; batimastat; benzodepa; bicalutamide; bisantrene hydrochloride; bisnafide
dimesylate; bizelesin; bleomycin sulfate; brequinar sodium; bropirimine; busulfan;
cactinomycin; calusterone; caracemide; carbetimer; carboplatin; carmustine; carubicin
hydrochloride; carzelesin; cedefingol; chlorambucil; cirolemycin; cisplatin; cladribine;
crisnatol mesylate; cyclophosphamide; cytarabine; dacarbazine; dactinomycin; daunorubicin
hydrochloride; decitabine; dexormaplatin; dezaguanine; dezaguanine mesylate; diaziquone;
docetaxel; doxorubicin; doxorubicin hydrochloride; droloxifene; droloxifene citrate;
dromostanolone propionate; duazomycin; edatrexate; eflornithine hydrochloride; elsamitrucin;
enloplatin; enpromate; epipropidine; epirubicin hydrochloride; erbulozole; esorubicin
hydrochloride; estramustine; estramustine phosphate sodium; etanidazole; etoposide;
etoposide phosphate; etoprine; fadrozole hydrochloride; fazarabine; fenretinide; floxuridine;
fludarabine phosphate; fluorouracil; flurocitabine; fosquidone; fostriecin sodium;
gemcitabine; gemcitabine hydrochloride; hydroxyurea; idarubicin hydrochloride; ifosfamide;
ilmofosine; interleukin II (including recombinant interleukin II, or rIL2), interferon
alfa-2a; interferon alfa-2b; interferon alfa-n1; interferon alfa-n3; interferon beta-Ia;
interferon gamma-Ib; iproplatin; irinotecan hydrochloride; lanreotide acetate; letrozole;
leuprolide acetate; liarozole hydrochloride; lometrexol sodium; lomustine; losoxantrone
hydrochloride; masoprocol; maytansine; mechlorethamine hydrochloride; megestrol acetate;
melengestrol acetate; melphalan; menogaril; mercaptopurine; methotrexate; methotrexate
sodium; metoprine; meturedepa; mitindomide; mitocarcin; mitocromin; mitogillin; mitomalcin;
mitomycin; mitosper; mitotane; mitoxantrone hydrochloride; mycophenolic acid; nocodazole;
nogalamycin; ormaplatin; oxisuran; paclitaxel; pegaspargase; peliomycin; pentamustine;
peplomycin sulfate; perfosfamide; pipobroman; piposulfan; piroxantrone hydrochloride;
plicamycin; plomestane; porfimer sodium; porfiromycin; prednimustine; procarbazine
hydrochloride; puromycin; puromycin hydrochloride; pyrazofurin; riboprine; rogletimide;
safingol; safingol hydrochloride; semustine; simtrazene; sparfosate sodium; sparsomycin;
spirogermanium hydrochloride; spiromustine; spiroplatin; streptonigrin; streptozocin;
sulofenur; talisomycin; tecogalan sodium; tegafur; teloxantrone hydrochloride; temoporfin;
teniposide; teroxirone; testolactone; thiamiprine; thioguanine; thiotepa; tiazofurin;
tirapazamine; toremifene citrate; trestolone acetate; triciribine phosphate; trimetrexate;
trimetrexate glucuronate; triptorelin; tubulozole hydrochloride; uracil mustard; uredepa;
vapreotide; verteporfin; vinblastine sulfate; vincristine sulfate; vindesine; vindesine
sulfate; vinepidine sulfate; vinglycinate sulfate; vinleurosine sulfate; vinorelbine
tartrate; vinrosidine sulfate; vinzolidine sulfate; vorozole; zeniplatin; zinostatin;
zorubicin hydrochloride. Other anti-cancer drugs include, but are not limited to:
20-epi-1,25 dihydroxyvitamin D3; 5-ethynyluracil; abiraterone; aclarubicin; acylfulvene;
adecypenol; adozelesin; aldesleukin; ALL-TK antagonists; altretamine; ambamustine;
amidox; amifostine; aminolevulinic acid; amrubicin; amsacrine; anagrelide; anastrozole;
andrographolide; angiogenesis inhibitors; antagonist D; antagonist G; antarelix; anti-dorsalizing
morphogenetic protein-1; antiandrogen, prostatic carcinoma; antiestrogen; antineoplaston;
antisense oligonucleotides; aphidicolin glycinate; apoptosis gene modulators; apoptosis
regulators; apurinic acid; ara-CDP-DL-PTBA; arginine deaminase; asulacrine; atamestane;
atrimustine; axinastatin 1; axinastatin 2; axinastatin 3; azasetron; azatoxin; azatyrosine;
baccatin III derivatives; balanol; batimastat; BCR/ABL antagonists; benzochlorins;
benzoylstaurosporine; beta lactam derivatives; beta-alethine; betaclamycin B; betulinic
acid; bFGF inhibitor; bicalutamide; bisantrene; bisaziridinylspermine; bisnafide;
bistratene A; bizelesin; breflate; bropirimine; budotitane; buthionine sulfoximine;
calcipotriol; calphostin C; camptothecin derivatives; canarypox IL-2; capecitabine;
carboxamide-amino-triazole; carboxyamidotriazole; CaRest M3; CARN 700; cartilage derived
inhibitor; carzelesin; casein kinase inhibitors (ICOS); castanospermine; cecropin
B; cetrorelix; chlorlns; chloroquinoxaline sulfonamide; cicaprost; cis-porphyrin;
cladribine; clomifene analogues; clotrimazole; collismycin A; collismycin B; combretastatin
A4; combretastatin analogue; conagenin; crambescidin 816; crisnatol; cryptophycin
8; cryptophycin A derivatives; curacin A; cyclopentanthraquinones; cycloplatam; cypemycin;
cytarabine ocfosfate; cytolytic factor; cytostatin; dacliximab; decitabine; dehydrodidemnin
B; deslorelin; dexamethasone; dexifosfamide; dexrazoxane; dexverapamil; diaziquone;
didemnin B; didox; diethylnorspermine; dihydro-5-azacytidine; dihydrotaxol, 9-; dioxamycin;
diphenyl spiromustine; docetaxel; docosanol; dolasetron; doxifluridine; droloxifene;
dronabinol; duocarmycin SA; ebselen; ecomustine; edelfosine; edrecolomab; eflornithine;
elemene; emitefur; epirubicin; epristeride; estramustine analogue; estrogen agonists;
estrogen antagonists; etanidazole; etoposide phosphate; exemestane; fadrozole; fazarabine;
fenretinide; filgrastim; finasteride; flavopiridol; flezelastine; fluasterone; fludarabine;
fluorodaunorunicin hydrochloride; forfenimex; formestane; fostriecin; fotemustine;
gadolinium texaphyrin; gallium nitrate; galocitabine; ganirelix; gelatinase inhibitors;
gemcitabine; glutathione inhibitors; hepsulfam; heregulin; hexamethylene bisacetamide;
hypericin; ibandronic acid; idarubicin; idoxifene; idramantone; ilmofosine; ilomastat;
imidazoacridones; imiquimod; immunostimulant peptides; insulin-like growth factor-1
receptor inhibitor; interferon agonists; interferons; interleukins; iobenguane; iododoxorubicin;
ipomeanol, 4-; iroplact; irsogladine; isobengazole; isohomohalicondrin B; itasetron;
jasplakinolide; kahalalide F; lamellarin-N triacetate; lanreotide; leinamycin; lenograstim;
lentinan sulfate; leptolstatin; letrozole; leukemia inhibiting factor; leukocyte alpha
interferon; leuprolide+estrogen+progesterone; leuprorelin; levamisole; liarozole;
linear polyamine analogue; lipophilic disaccharide peptide; lipophilic platinum compounds;
lissoclinamide 7; lobaplatin; lombricine; lometrexol; lonidamine; losoxantrone; lovastatin;
loxoribine; lurtotecan; lutetium texaphyrin; lysofylline; lytic peptides; maitansine;
mannostatin A; marimastat; masoprocol; maspin; matrilysin inhibitors; matrix metalloproteinase
inhibitors; menogaril; merbarone; meterelin; methioninase; metoclopramide; MIF inhibitor;
mifepristone; miltefosine; mirimostim; mismatched double stranded RNA; mitoguazone;
mitolactol; mitomycin analogues; mitonafide; mitotoxin fibroblasts growth factor-saporin;
mitoxantrone; mofarotene; molgramostim; monoclonal antibody, human chorionic gonadotrophin;
monophosphoryl lipid A+myobacterium cell wall sk; mopidamol; multiple drug resistance
gene inhibitor; multiple tumor suppressor 1-based therapy; mustard anticancer agent;
mycaperoxide B; mycobacterial cell wall extract; myriaporone; N-acetyldinaline; N-substituted
benzamides; nafarelin; nagrestip; naloxone+pentazocine; napavin; naphterpin; nartograstim;
nedaplatin; nemorubicin; neridronic acid; neutral endopeptidase; nilutamide; nisamycin;
nitric oxide modulators; nitroxide antioxidant; nitrullyn; 06-benzylguanine; octreotide;
okicenone; oligonucleotides; onapristone; ondansetron; ondansetron; oracin; oral cytokine
inducer; ormaplatin; osaterone; oxaliplatin; oxaunomycin; paclitaxel; paclitaxel analogues;
paclitaxel derivatives; palauamine; palmitoylrhizoxin; pamidronic acid; panaxytriol;
panomifene; parabactin; pazelliptine; pegaspargase; peldesine; pentosan polysulfate
sodium; pentostatin; pentrozole; perflubron; perfosfamide; perillyl alcohol; phenazinomycin;
phenylacetate; phosphatase inhibitors; picibanil; pilocarpine hydrochloride; pirarubicin;
piritrexim; placetin A; placetin B; plasminogen activator inhibitor; platinum complex;
platinum compounds; platinum-triamine complex; porfimer sodium; porfiromycin; prednisone;
propyl bis-acridone; prostaglandin J2; proteasome inhibitors; protein A-based immune
modulator; protein kinase C inhibitor; protein kinase C inhibitors, microalgal; protein
tyrosine phosphatase inhibitors; purine nucleoside phosphorylase inhibitors; purpurins;
pyrazoloacridine; pyridoxylated hemoglobin polyoxyethylene conjugate; raf antagonists;
raltitrexed; ramosetron; ras farnesyl protein transferase inhibitors; ras inhibitors;
ras-GAP inhibitor; retelliptine demethylated; rhenium Re 186 etidronate; rhizoxin;
ribozymes; RII retinamide; rogletimide; rohitukine; romurtide; roquinimex; rubiginone
B1; ruboxyl; safingol; saintopin; SarCNU; sarcophytol A; sargramostim; Sdi 1 mimetics;
semustine; senescence derived inhibitor 1; sense oligonucleotides; signal transduction
inhibitors; signal transduction modulators; single chain antigen binding protein;
sizofiran; sobuzoxane; sodium borocaptate; sodium phenylacetate; solverol; somatomedin
binding protein; sonermin; sparfosic acid; spicamycin D; spiromustine; splenopentin;
spongistatin 1; squalamine; stem cell inhibitor; stem-cell division inhibitors; stipiamide;
stromelysin inhibitors; sulfinosine; superactive vasoactive intestinal peptide antagonist;
suradista; suramin; swainsonine; synthetic glycosaminoglycans; tallimustine; tamoxifen
methiodide; tauromustine; tazarotene; tecogalan sodium; tegafur; tellurapyrylium;
telomerase inhibitors; temoporfin; temozolomide; teniposide; tetrachlorodecaoxide;
tetrazomine; thaliblastine; thiocoraline; thrombopoietin; thrombopoietin mimetic;
thymalfasin; thymopoietin receptor agonist; thymotrinan; thyroid stimulating hormone;
tin ethyl etiopurpurin; tirapazamine; titanocene bichloride; topsentin; toremifene;
totipotent stem cell factor; translation inhibitors; tretinoin; triacetyluridine;
triciribine; trimetrexate; triptorelin; tropisetron; turosteride; tyrosine kinase
inhibitors; tyrphostins; UBC inhibitors; ubenimex; urogenital sinus-derived growth
inhibitory factor; urokinase receptor antagonists; vapreotide; variolin B; vector
system, erythrocyte gene therapy; velaresol; veramine; verdins; verteporfin; vinorelbine;
vinxaltine; vitaxin; vorozole; zanoterone; zeniplatin; zilascorb; and zinostatin stimalamer.
In one embodiment, the anti-cancer drug is 5-fluorouracil, taxol, or leucovorin.
Methods Related to Treatment of Conditions Involving Microorganisms
[0217] Disclosed is a method for prophylactically treating a human or a mammal against infection
by a microorganism, comprising administering to a human or mammal an effective amount
of one or more of the disclosed H1F-1α prolyl hydroxylase inhibitors.
[0218] Further disclosed is a method for decreasing the virulence of a microorganism when
a human or a mammal is infected with a microorganism, comprising administering to
a human or mammal an effective amount of one or more of the disclosed HIF-1α prolyl
hydroxylase inhibitors.
[0219] Yet further disclosed is a method for treating an infection in a human or mammal
caused by a microorganism, comprising administering to a human or mammal an effective
amount of one or more of the disclosed HIF-1α prolyl hydroxylase inhibitors.
[0220] Still further disclosed is a method for treating a human or mammal diagnosed with
an infection caused by a microorganism, comprising administering to a human or mammal
an effective amount of one or more of the disclosed HIF-1α prolyl hydroxylase inhibitors.
[0221] Also disclosed is a method for preventing transmission of a disease caused by a microorganism
from a human or mammal to a human or mammal, comprising administering to a human or
mammal an effective amount of one or more of the disclosed HIF-1α prolyl hydroxylase
inhibitors.
[0222] Still yet further disclosed is a method for preventing infection of a human or a
mammal during a surgical procedure, comprising administering to a human or mammal
an effective amount of one or more of the disclosed HIF-1α prolyl hydroxylase inhibitors.
[0223] The microorganism can be any benign or virulent microorganism, for example, bacteria,
viruses, yeasts, fungi, or parasites. The following are non-limiting examples of microorganisms
that can be affected by the disclosed HIF-1α prolyl hydroxylase inhibitors. By the
term "affected" is meant, the virulence of the microorganism is reduced, diminished
or eliminated. The cause of the reduction, diminishment, or elimination of the virulence
can be from the stabilization of HIF-1 and/or from the increased level of phagocytosis
due to the administration of one or more of the disclosed HIF-1α prolyl hydroxylase
inhibitors.
[0224] Acinetobacter calcoaceticus, Acinetobacter- haemolyticus, Aeromonas hydrophilia, Agrobacterium
tumefaciens, Bacillus anthracis, Bacillus halodurans, Bacillus subtilis, Bacteroides
distasonis, Bacteroides eggerthii, Bacteroides fragilis, Bacteroides ovalus, Bacteroides
3452A homology group, Bacteroides splanchnicus, Bacteroides thetaiotaomicron, Bacteroides
uniformis, Bacteroides vulgatus, Bordetella bronchiseptica, Bordetella parapertussis,
Bordetella pertussis, Borrelia burgdorferi, Branhamella catarrhalis, Brucella melitensis,
Burkholderia cepacia, Burkholderia pseudomallei, Campylobacter coli, Campylobacterfetus,
Campylobacter jejuni, Caulobacter crescentus, Citrobacter freundii, Clostridium difficile,
Clostridium perfingens, Corynebacterium diphtheriae, Corynebacterium glutamicum, Corynebacterium
ulcerans, Edwardsiella tarda, Enterobacter aerogenes, Erwinia chrysanthemi, Enterobacter
cloacae, Enterococcus faecalis, Enterococcus faecium, Escherichia coli, Fruncisella
tularensis, Gardnerella vaginalis, Haemophilus ducreyi, Haemophilus haemolyticus,
Haemophilus influenzae, Haemophilus parahaemolyticus, Haemophilus parainfluenzae,
Helicobacter pylori, Klebsiella oxytoca, Klebsiella pneumoniae, Kluyvera cryocrescens,
Legionella pneumophila, Listeria innocua, Listeria monocytogenes, Listeria welshimeri,
Methanosarcina acetivorans, Methanosarcina mazei, Morganella morganii, Mycobacterium
avium, Mycobacterium intracellulare, Mycobacterium leprae, Mycobacterium tuberculosis,
Mesorhizobium loti, Neisseria gonorrhoeae, Neisseria meningitidis, Pasteurella haemolytica,
Pasteurella multocida, Providencia alcalifaciens, Providencia rettgeri, Providencia
stuartii, Proteus mirubilis, Proteus vulgaris, Pseudomonas acidovorans, Pseudomonas
aeruginosa, Pseudomonas alcaligenes, Pseudomonas fluorescens, Pseudomonas putida,
Ralstonia solanacearum, Salmonella enterica subsp. enteridtidis, Salmonella enterica
subsp. paratyphi, Salnionella enterica, subsp. typhimurium, Salmonella enterica, subsp.
typhi, Serratia marcescens, Shigella dysenteriae, Shigella flexneri, Shigella sonnei,
Sinorhizobium meliloti, Staphylococcus aureus, Streptococcus criceti, Staphylococcus
epidemmidis, Staphylococcus haemolyticus, Staphylococcus hominis, Staphylococcus hyicus,
Staphylococcus intermedius, Stenotrophomonas maltophilia, Staphylococcus saccharolyticus,
Staphylococcus saprophyticus, Staphylococcus sciuri, Streptomyces avermitilis, Streptomyces
coelicolor, Streptococcus agalactiae, Streptococcus pneumoniae, Streptococcus pyogenes
Sulfobalblobus soffiataricus, Thermotoga maritima, Vibrio cholerae, Vibrio parahaemoliyticus,
Yogesella indigofera, Xanthomonas axonopodis, Xanthomonas campestris, Yersinia enterocolitica,
Yersinia intermedia, Yersinia pestis, and Yersinia pseudotuberculosis
Methods Relating to Vaccination or Innoculation
[0225] Disclosed herein are methods for enhancing the effectiveness of a vaccine, comprising
co-administering to a human or mammal a vaccine in combination with one or more HIF-1α
prolyl hydroxylase inhibitors.
[0226] Non-limiting examples of vaccines are those for stimulating antibodies against hepatitis,
influenza, measles, rubella, tetanus, polio, rabies, and the like.
[0227] Therefore, the disclosed methods includes administering, or in the case of contacting
cells
in vitro, in vivo or
ex vivo, the one or more HIF-1α prolyl hydroxylase inhibitors and any co-administered compounds
topically, buccally, orally, intradermally, subcutaneously, mucosally in the eye,
vagina, rectum, and nose, intravenously, and intramuscularly.
PROCEDURES
EGLN-1 activity assay
[0228] The EGLN-1 (or EGLN-3) enzyme activity is determined using mass spectrometry (matrix-assisted
laser desorption ionization, time-of-flight MS, MALDI-TOF MS. Recombinant human EGLN-1-179/426
is prepared as described above and in the Supplemental Data. Full-length recombinant
human EGLN-3 is prepared in a similar way, however it is necessary to use the His-MBP-TVMV-EGLN-3
fusion for the assay due to the instability of the cleaved protein. For both enzymes,
the HIF-1α peptide corresponding to residues 556-574 is used as substrate. The reaction
is conducted in a total volume of 50 µL containing TrisCl (5 mM, pH 7.5), ascorbate
(120 µM), 2-oxoglutarate (3.2 µM), HIF-1α (8.6 µM), and bovine serum albumin (0.01%).
The enzyme, quantity predetermined to hydroxylate 20% of substrate in 20 minutes,
is added to start the reaction. Where inhibitors are used, compounds are prepared
in dimethyl sulfoxide at 10-fold final assay concentration. After 20 minutes at room
temperature, the reaction is stopped by transferring 10 µL of reaction mixture to
50 µL of a mass spectrometry matrix solution (α-cyano-4-hydroxycinnamic acid, 5 mg/mL
in 50% acetonitrile/0.1% TFA, 5 mM NH
4PO
4). Two microliters of the mixture is spotted onto a MALDI-TOF MS target plate for
analysis with an Applied Biosystems (Foster City, CA) 4700 Proteomics Analyzer MALDI-TOF
MS equipped with a Nd:YAG laser (355 nm, 3 ns pulse width, 200 Hz repetition rate).
Hydroxylated peptide product is identified from substrate by the gain of 16 Da. Data
defined as percent conversion of substrate to product is analyzed in GraphPad Prism
4 to calculate IC
50 values.
VEGF ELISA assay
[0229] HEK293 cells are seeded in 96-well poly-lysine coated plates at 20,000 cells per
well in DMEM (10% FBS, 1% NEAA, 0.1% glutamine). Following overnight incubation, the
cells are washed with 100 µL of Opti-MEM (Gibco, Carlsbad, CA) to remove serum. Compound
in DMSO is serially diluted (beginning with 100 µM) in Opti-MEM and added to the cells.
The conditioned media is analyzed for VEGF with a Quantikine human VEGF immunoassay
kit (R&D Systems, Minneapolis, MN). Optical density measurements at 450 nm are recorded
using the Spectra Max 250 (Molecular Devices, Sunnyvale, CA). Data defined as % of
DFO stimulation is used to calculate EC
50 values with GraphPad Prism 4 software (San Diego, CA).
Mouse Ischemic Hindlimb Study
[0230] All animal work is conducted in accordance with the Guide for the Care and Use of
Laboratory Animals (National Academy of Sciences; Copyright ©1996). Used in these
experiments were 9-10 week old male C57B1/6 mice from Charles River Laboratory (Portage,
MI). The mice are orally dosed with vehicle (aqueous carbonate buffer, 50 mM; pH 9.0)
or with the compound to be tested in vehicle at 50 mg/kg or 100 mg/kg. The animals
are dosed three times: day 1 at 8 AM and 5 PM, and on day 2 at 8 AM. One hour after
the first dose, unilateral arterial ligation is performed under anesthesia using isoflurane.
The femoral artery is ligated proximal to the origin of the popliteal artery. The
contralateral limb undergoes a sham surgical procedure. Ligation is performed in an
alternating fashion between right and left hindlimbs. Two hours after the 8 AM dosing
on day 2, blood is obtained by ventricular stick while the mice are anesthetized with
isoflurane. Serum samples for EPO analysis are obtained using gel clot serum separation
tubes. Heart, liver, and gastrocnemius muscles are harvested, snap-frozen in liquid
nitrogen, and stored in -80°C until use.
Mouse Serum EPO Assay
[0231] The mouse serum EPO is detected using Mouse Quantikine Erythropoietin ELISA kit from
R&D Systems according to manufacturer's instructions.
Mouse Tissue HIF Western Blot Analysis
[0232] Tissues from mice stored at -80°C are powdered with mortar and pestle chilled with
liquid nitrogen. Nuclear extracts are prepared using an NE-PER kit (Pierce Biotechnology).
For immunoprecipitation, nuclear extract is added to monoclonal antibody to HIF-1α
(Novus, Littleton, CO) at a tissue to antibody ratio of 200:1. The suspension is incubated
in a conical micro centrifuge tube for 4 hours at 4 °C. Protein A/G-coupled agarose
beads (40 µL of a 50% suspension) are then added to the tube. Following overnight
tumbling at 4 °C, the beads are washed 3 times with ice-cold phosphate buffered saline.
The beads are then prepared for SDS-PAGE with 40 µL of Laemmli sample buffer. Proteins
separated on SDS-PAGE are transferred onto nitrocellulose sheets with XCell-II Blot
Module system (Invitrogen, Carlsbad, CA). The blots are blocked with 5% BSA prior
to incubation with a rabbit antibody to HIF-1α at 1:100 dilution (Novus). The blots
are then washed with Tris-buffered saline/Tween-20 buffer and incubated with horseradish
peroxidase-conjugated goat anti-rabbit secondary antibody (Pierce, Rockford, IL).
Blots are developed with the ECL reagent (Amersham, Piscataway, NJ). Images of blots
are captured with an Epson Expression 1600 scanner.
[0234] Compound F2 was further tested in the mouse serum EPO assay described herein above
and found to have an EPO EC
50 = 14 µM.
Enhanced Neutrophil Activity
[0236] For bacterial assays,
S.
Aureus (ATCC 33591) can be grown in Todd-Hewitt broth (THB) to logarithmic phase (OD
600 of 0.4 or ∼5 x 10
7 cfu/mL) and then pelleted, washed, and resuspended in PBS or RPMI 1640 tissue-culture
medium to the desired concentration. Venous blood from healthy volunteers can be used
for whole blood and neutrophil isolation. Neutrophils can be purified using the PolyMorphPrep
Kit (Axis Shield) in accordance with manufacturer's instructions. Human monocytic
cell line U937 can be propagated in RPMI 1640 plus 10% fetal calf serum, 1 mmol/L
NaPyr, 10 mmol/L HEPES, and glucose. Whole blood or phagocytic cells can be preincubated
with mimosine (Sigma-Aldrich) (0-500 µmol/L) for 2-4 hours then challenged with
S. Aureus (either 10
5 cfu in 100 µL added to 300 µL of whole blood or at an MOI of 1 bacterium/cell for
isolated phagocytes). Aliquots are then plated on THB agar after 30 (whole blood and
neutrophils) or 60 (U937 monocytes) min for enumeration of surviving
S.
Aureus colony-forming units.
EXAMPLE 5
[0237] Isolated human neutrophils were pre-incubated for 1 hour at 37 °C with a control
consisting of dimethyl sulfoxide (DMSO), 50 µM and 200 µM of a compound disclosed
in Table VIII.
Staphylococcus aureus (Newman strain) was then added to the neutrophils at an MOI of approximately 0.1
(1 bacterium for every 10 neutrophils). Samples were taken at 60 and 90 minutes wherein
the neutrophils were lysed with water, and the total bacteria remaining were enumerated
on Todd-Hewitt broth (THB) agar plates.
[0238] Figure 2 depicts the effectiveness of a compound disclosed in Table VIII in providing enhanced
killing of
S. aureus (Newman strain) at concentrations of 50 µM and 200 µM versus control. As can be seen
in
Figure 2, at 90 minutes post-infection, approximately half of the colony forming units are
absent at a concentration of 200 µM.
EXAMPLE 6
[0239] Cells from the human monocyte cell line U937 were pre-incubated for 2 hours at 37
°C under an atmosphere of 5% CO
2 with a control consisting of DMSO and 10 µM of a compound disclosed in Table VIII.
Staphylococcus aureus (virulent Newman strain) was then added to the cells at an MOI of approximately 1
(1 bacterium for every 1 cell). Samples are drawn at 30, 60, 90 and 120 minutes post-infection.
The U937 cells were lysed with Triton
™, and the amount of bacteria remaining were enumerated on THB agar plates.
[0240] As depicted in
Figure 3, 4-prolyl hydroxylase inhibitor a compound disclosed in Table VIII is effective in
killing
S.
aureus when compared to a control (DMSO). At 120 minutes, a compound disclosed in Table
VIII produces an 84% kill of Newman strain
S. aureus when the monocyte cells are treated with 10 µM of a compound disclosed in Table VIII,
thereby showing increased phagocytosis due to extended neutrophil life span.
EXAMPLE 7
[0241] Two samples of cells from the human monocyte cell line U937 were pre-treated with
10 µM of a compound disclosed in Table VIII. One sample was pre-incubated for 1 hour
and the other sample pre-incubated for 2 hours, both at 37 °C under an atmosphere
of 5% CO
2.
S.
aureus (virulent Newman strain) was then added to the cells at an MOI of approximately 1-2
(1-2 bacteria for every 1 cell). Aliquots of cells were removed from each sample at
30, 60, 90, and 120 minutes post-infection, the U937 cells were immediately lysed
with Triton
™, and total of remaining bacteria remaining were enumerated on THB agar plates.
[0242] As depicted in
Figure 4, U937 monocyte cells pre-treated with 10 µM of a compound disclosed in Table VIII
for 1 hour (black bars) had almost no colony forming units present 120 minutes post-infection,
whereas the cells pre-treated two hours prior to infection had approximately 15% colony
forming units present as compared to cells that were untreated. In addition,
Figure 4 indicates that within 1 hour after the U937 monocyte cells had bee exposed to
S.
aureus (Newman strain), the number of colony forming units present was significantly reduced
relative to cells receiving no HIF-1α inhibitor.
EXAMPLE 8
[0243] Two samples of cells from the human monocyte cell line U937 were pre-treated with
10 µM of a compound disclosed in Table VIII for 1 at 37 °C under an atmosphere of
5% CO
2.
S.
aureus (Newman strain) was added to one sample and to the other was added methicillin-resistant
S.
aureus (MRSA). Both bacteria were added at an MOI of approximately 2-3 (2-3 bacteria for
every 1 cell). Aliquots of cells were removed from each sample at 30, 60, 90, and
120 minutes post-infection. The U937 cells were immediately lysed with Triton
™, and total of remaining bacteria remaining were enumerated on THB agar plates.
[0244] As depicted in
Figure 5, at120 minutes post-infection, the MRSA infected cells had only 25% of the average
percentage of colony forming units present when compared to control as represented
by the black bars. Also depicted in
Figure 5, at 60 minutes post-infection, the Newman strain of
S. aureus had only approximately 12% of the average percentage of colony forming units present
when compared to control, and almost no colony forming units present at 120 minutes
post-infection as represented by the hatched bars.
EXAMPLE 9
[0245] Two samples of cells from the human monocyte cell line U937 treated with 10 µM a
compound disclosed in Table VIII were infected with either
S.
aureus (Newman strain) and methicillin-resistant
S.
aureus (MRSA). Both bacteria were added at an MOI of approximately 2-3 (2-3 bacteria for
every 1 cell). Aliquots of cells were removed from each sample at 30, 60, 90, and
120 minutes post-infection. The U937 cells were immediately lysed with Triton
™, and total remaining bacteria were enumerated on THB agar plates.
[0246] As depicted in
Figure 6, even without pre-treatment with a compound disclosed in Table VIII, at 60 minutes
post-infection, the Newman strain of
S.
aureus had only 25% of the average percentage of colony forming units present when compared
to control as represented by the black bars. The MRSA strain was reduced to less than
approximately 40% of the average percentage of colony forming units present when compared
to control as represented by the hatched bars.
EXAMPLE 10
[0247] Three samples of cells from the human monocyte cell line U937 were treated with 100
µM mimosine, 2 µg/mL vancomycin or 10 µM of a compound disclosed in Table VIII. Each
sample was infected with either
S.
aureus (Newman strain) or methicillin-resistant
S.
aureus (MRSA). Both bacteria were added at an MOI of approximately 2-3 (2-3 bacteria for
every 1 cell). At 120 minutes post-infection aliquots were withdrawn from all six
samples and the U937 cells were immediately lysed with Triton
™, and total remaining bacteria were enumerated on THB agar plates.
[0248] As depicted in
Figure 7, 10 µM a compound disclosed in Table VIII enhanced kill of both bacterial strains,
i.e., S. aureus, Newman (hatched bars) or MRSA (black bars), when compared to mimosine treated cells.
Refering to the hatched bars representing Newman strain, as further depicted in
Figure 7, the sample treated with 10 µM a compound disclosed in Table VIII had a lower average
percentage of colony forming units present than the cells treated with vancomycin.
The U937 cells infected with MRSA (black bars) had approximately 40% of the colony
forming units present versus untreated cells and less than half the number of those
treated with mimosine.
[0249] Figure 8 depicts the average percentage of colony forming units present (Newman strain) versus
control for human monocyte cells (U937) at 30, 60, 90, and 120 minutes post-infection,
when treated with 10 µM a compound disclosed in Table VIII. The black bars represent
treatment with a compound disclosed in Table VIII beginning at the time of infection
with
S.
aureus, the hatched bars represent cells pretreated with a compound disclosed in Table VIII
and white bars represent cells pretreated two hours prior to infection with
S. aureus.
[0250] Figure 9 depicts the average percent of colony forming units present at 120 minutes post-infection
vs DMSO (control) when HaCaT cells are pre-treated for 1 hour according to the examples
above with 800 µM mimosine, 10 µM of compound disclosed in Table VIII or 1 µg mL vancomycin
followed by inoculation with
S.
aureus (Newman strain, hatched bars) and methicillin-resistant
S.
aureus (MRSA, black bars).
Figure 10 depicts the average percent of colony forming units present at 30, 60, 90, and 120
minutes post-infection for Newman strain of
S. aureus (hatched bars) and MRSA (black bars) when HaCaT cells are pre-treated for 1 hour
according to the examples above with 10 µM of a compound disclosed in Table VIII.
[0251] Figure 11 depicts the up regulation of phosphoglycerate kinase (PGK) expression in wild type
murine embryonic fibroblasts as a result of treatment with compound a
compound disclosed in Table VIII at dosages of 1 µM (E), 10 µM (F), and 50 µM (G) vs. wild type control (H) and the
lack of up regulation of PGK expression in HIF-1 knock out cells as a result of treatment
with a compound disclosed in Table VIII at dosages of 1 µM (A), 10 µM (B), and 50
µM (C) and HIF-1 knock out control (D). Both cell types were treated for 7 hours.
[0252] Figure 12 depicts the up regulation of phosphoglycerate kinase (PGK) expression in wild type
murine embryonic fibroblasts as a result of treatment with a compound disclosed in
Table VIII at dosages of 1 µM (E), 10 µM (F), vs. wild type control (G) and the lack
of up regulation of PGK expression in HIF-1 knock out cells as a result of treatment
with a compound disclosed in Table VIII at dosages of 1 µM (A), 10 µM (B), and 50
µM (C) and HIF-1 knock out control (D).
[0253] Figure 13 depicts the up regulation of phosphoglycerate kinase (PGK) expression in wild type
murine embryonic fibroblasts as a result of treatment with a compound disclosed in
Table VIII at dosages of 1 µM (E), 10 µM (F), and 50 µM (G) vs. wild type control
(H) and the lack of up regulation of PGK expression in HIF-1 knock out cells as a
result of treatment with a compound disclosed in Table VIII at dosages of 1 µM (A),
10 µM (B), and 50 µM (C) and HIF-1 knock out control (D).
[0254] Vascular Endothelial Growth Factor (VEGF) is dependent upon the presence of HIF-1
in cells.
Figure 14 depicts the up regulation of vascular endothelia growth factor (VEGF) expression
in wild type murine embryonic fibroblasts as a result of treatment with a compound
disclosed in Table VIII at dosages of 1 µM (E), 10 µM (F), and 50 µM (G) vs. control
(H) and the lack of up regulation of VEGF expression in HIF-1 knock out cells treated
with compound a compound disclosed in Table VIII at dosages of 1 µM (A), 10 µM (B),
and 50 µM (C) and HIF-1 knock out control (D). Both cell types were treated for 7
hours. As seen in
Figure 14, VEGF is increased when dosed at 10 µM (F) and 50 µM (G). In HIF-1 knock out cells,
there is no increase in PGK up regulation when HIF-1 knock out cells are dosed at
1 µM (A), 10 µM (B), and 50 µM (C) when compared to wild type control (H) and HIF-1
knock out control (D).
Wound Healing
EXAMPLE 11
[0255] Twenty-four (24) mice were divided into three groups. Group 2 animals were administered
bacterial inoculum (
Staphylococcus aureus antibiotic sensitive Newman strain [ATCC #25904]) by subcutaneous injection on Day
0 and received 10 µM of a compound disclosed in Table VIII for 6 days starting at
2 hours post-infection (Days 0-5). Group 1 received subcutaneous injections of DMSO.
Group 3 served as a control group and received no treatment. Lesion size was monitored
daily during the study. Only open wounds were considered lesions; bumps and white
patches without an open wound were not measured for lesion size. On Day 7, the final
lesion size was measured and mice were sacrificed for determination of bacterial load
in skin and kidney. Day 7 post-infection, mice were sacrificed after final lesion
size measurement and the lesioned skin tissue and both kidneys were collected. Skin
and kidneys were homogenized in phosphate buffered saline, serially diluted, and plated
on Todd-Hewitt agar plates to enumerate colony forming units of bacteria.
[0256] Figure 15 shows the significant reduction in the size of skin lesions (wounds) for animals
in Group 1 (solid circles (●)) treated with 10µM of a compound disclosed in Table
VIII versus animal treated with DMSO (solid squares (■)). As depicted in
Figure 15, mice infected with Newman strain of S. aureus followed by treatment with 10 µM of
a compound disclosed in Table VIII or DMSO (control) at 2 hours post-infection. The
data show the statistically significant reduction in the size of skin lesions (wounds)
for animals treated with a compound disclosed in Table VIII (solid circles (●)) or
DMSO (solid squares (■)).
[0257] Figure 16 shows the significant reduction in the size of skin lesions (wounds) for animals
in Group 1 (solid circles (●)) treated with 10µM of a compound disclosed in Table
VIII versus untreated animals (solid triangles (▲)). As depicted in Figure 16, mice
infected with Newman strain of S. aureus followed by treatment with 10 µM of a compound
disclosed in Table VIII or no treatment at 2 hours post-infection. The data show the
reduction in the size of skin lesions (wounds) for animals treated with a compound
disclosed in Table VIII (solid circles (●)) or untreated (solid triangles (▲)).
EXAMPLE 12
[0258] Twenty-four (24) mice were divided into three groups. Group 1 animals were administered
bacterial inoculum (
Staphylococcus aureus antibiotic sensitive Newman strain [ATCC #25904]) by subcutaneous injection on Day
0 and received 10 µM of a compound disclosed in Table VIII for 6 days starting at
2 hours post-infection (Days 0-5). Group 2 received subcutaneous injections of DMSO.
Group 3 served as a control group and received no treatment. Lesion size was monitored
daily during the study. Only open wounds were considered lesions; bumps and white
patches without an open wound were not measured for lesion size. Day 7 post-infection,
mice were sacrificed after final lesion size measurement and lesioned skin tissue
and both kidneys were collected. Skin and kidneys were homogenized in phosphate buffered
saline, serially diluted, and plated on Todd-Hewitt agar plates to enumerate colony
forming units of bacteria.
[0259] Figure 17 is a plot histogram wherein the number of observed colony forming units per gram
of skin tissue is depicted. The straight lines indicate the mean value for each group.
The results for the untreated group are plotted under (A), the results for the group
treated with DMSO are plotted under (B) and results for the group treated with 10
µM of a compound disclosed in Table VIII are plotted under (C).
[0260] Figure 18 is a plot of the observed colony forming units of bacteria found in the kidneys of
the animals. The results for the untreated group are plotted under (A), the results
for the group treated with DMSO are plotted under (B) and results for the group treated
with 10 µM of a compound disclosed in Table VIII are plotted under (C). As can be
seen from these data, half of the animals treated with the HIF-1α prolyl hydroxylase
inhibitor disclosed in Table VIII had no bacteria in the kidney indicating that the
compound disclosed in Table VIII was able to systemically prevent spread of the infection
from the wound to the kidney.
EXAMPLE 13
[0261] Twenty (20) mice were divided into two groups. Group 1 animals were administered
bacterial inoculum (
Streptococcus pyogenes NZ131 [M49 strain]) by subcutaneous injection on Day 0 and were pretreated with compound
a compound disclosed in Table VIII once per day for 4 days, starting 2 hours pre-infection
(Days 0-3). Compound a compound disclosed in Table VIII was formulated in cyclodextran
and diluted in distilled water prior to subcutaneous injection, at a dose of 0.5 mg/kg.
Lesion size was monitored daily during the study. Only open wounds were considered
lesions; bumps and white patches without an open wound were not measured for lesion
size. On Day 4 post-infection, mice were sacrificed after final lesion size measurement
and lesioned skin tissue and both kidneys were collected. Skin and kidneys were homogenized
in phosphate buffered saline, serially diluted, and plated on Todd-Hewitt agar plates
to enumerate colony forming units of bacteria.
[0262] Figure 19 depicts the results of Example 13 wherein 2 groups of animals are treated with
Streptococcus pyogenes NZ131 [M49 strain]. The data show the reduction in the size of skin lesions (wounds)
for animals in Group 1 (solid triangles (A)) treated with 0.5 mg/kg of a compound
disclosed in Table VIII versus animal treated with vehicle control (cyclodextran)
(solid circles (●)),
Figure 20 is a plot histogram that also depicts the results of Example 12 wherein the number
of colony forming units for the observed skin lesions on animals treated with vehicle
control (cyclodextran) are plotted under (A) and results for the group treated with
0.5 mg/kg of a compound disclosed in Table VIII are plotted under (B).
KITS
[0263] Also disclosed are kits comprising the HIF-1α prolyl hydroxylase inhibitors be delivered
into a human, mammal, or cell. The kits can comprise one or more packaged unit doses
of a composition comprising one or more HIF-1α prolyl hydroxylase inhibitors to be
delivered into a human, mammal, or cell. The units dosage ampules or multidose containers,
in which the HIF-1α prolyl hydroxylase inhibitors to be delivered are packaged prior
to use, can comprise an hermetically sealed container enclosing an amount of polynucleotide
or solution containing a substance suitable for a pharmaceutically effective dose
thereof, or multiples of an effective dose. The HIF-1α prolyl hydroxylase inhibitor
can be packaged as a sterile formulation, and the hermetically sealed container is
designed to preserve sterility of the formulation until use.
[0264] The disclosed HIF-1α prolyl hydroxylase inhibitors can also be present in liquids,
emulsions, or suspensions for delivery of active therapeutic agents in aerosol form
to cavities of the body such as the nose, throat, or bronchial passages. The ratio
of HIF-1α prolyl hydroxylase inhibitors to the other compounding agents in these preparations
will vary as the dosage form requires.
[0265] Depending on the intended mode of administration, the pharmaceutical compositions
can be in the form of solid, semi-solid or liquid dosage forms, such as, for example,
tablets, suppositories, pills, capsules, powders, liquids, suspensions, lotions, creams,
gels, or the like, preferably in unit dosage form suitable for single administration
of a precise dosage. The compositions will include, as noted above, an effective amount
of the HIF-1α prolyl hydroxylase inhibitor in combination with a pharmaceutically
acceptable carrier and, in addition, can include other medicinal agents, pharmaceutical
agents, carriers, adjuvants, diluents, etc.
[0266] For solid compositions, conventional nontoxic solid carriers include, for example,
pharmaceutical grades of mannitol, lactose, starch, magnesium stearate, sodium saccharin,
talc, cellulose, glucose, sucrose, magnesium carbonate, and the like. Liquid pharmaceutically
administrable compositions can, for example, be prepared by dissolving, dispersing,
etc., an active compound as described herein and optional pharmaceutical adjuvants
in an excipient, such as, for example, water, saline aqueous dextrose, glycerol, ethanol,
and the like, to thereby form a solution or suspension. If desired, the pharmaceutical
composition to be administered can also contain minor amounts of nontoxic auxiliary
substances such as wetting or emulsifying agents, pH buffering agents and the like,
for example, sodium acetate, sorbitan monolaurate, triethanolamine sodium acetate,
triethanolamine oleate, etc. Actual methods of preparing such dosage forms are known,
or will be apparent, to those skilled in this art; for example see
Remington's Pharmaceutical Sciences, referenced above.
[0267] Parental administration, if used, is generally characterized by injection. Injectables
can be prepared in conventional forms, either as liquid solutions or suspensions,
solid forms suitable for solution or suspension in liquid prior to injection, or as
emulsions. A more recently revised approach for parental administration involves use
of a slow release or sustained release system, such that a constant level of dosage
is maintained.
See, e.g., U.S. Patent No. 3,710,795.
[0268] When the HIF-1α prolyl hydroxylase inhibitors are to be delivered into a mammal other
than a human, the mammal can be a non-human primate, horse, pig, rabbit, dog, sheep,
goat, cow, cat, guinea pig or rodent. The terms human and mammal do not denote a particular
age or sex. Thus, adult and newborn subjects, as well as fetuses, whether male or
female, are intended to be covered. A patient, subject, human or mammal refers to
a subject afflicted with a disease or disorder. The term "patient" includes human
and veterinary subjects.
[0269] While particular embodiments of the present disclosure have been illustrated and
described, it would be obvious to those skilled in the art that various other changes
and modifications can be made without departing from the scope of the disclosure.
It is therefore intended to cover in the appended claims all such changes and modifications
that are within the scope of this disclosure.